Genes associated with schizophrenia adhd and bipolar disorders

ABSTRACT

Disclosed are methods for diagnosing, monitoring the progression of, and treating schizophrenia, bipolar disorder, and/or ADHD based upon genes that are differentially expressed in said disorders at baseline, or at different timepoints following an acute stress exposure. Also disclosed are methods for identifying agents useful in the treatment of schizophrenia, bipolar disorder, and/or ADHD, methods for monitoring the efficacy of a treatment for schizophrenia, bipolar disorder, and/or ADHD, methods for preventing and treating schizophrenia, bipolar disorder, and/or ADHD, and an animal model for schizophrenia, bipolar disorder, and/or ADHD.

FIELD OF THE INVENTION

The present invention relates generally to the field of neurological andphysiological dysfunctions associated with schizophrenia,attention-deficit-hyperactivity disorder (ADHD) and bipolar disorder.The invention further relates to genes which, when varied in theirnormal expression pattern, are associated with schizophrenia, ADHD andbipolar disorder. Thus, the present invention relates to the novel useof known genes in schizophrenia, ADHD and bipolar disorder. The presentinvention also relates to methods for diagnosing and observing diseaseprogression of schizophrenia, ADHD and bipolar disorder. The presentinvention further relates to methods for identifying agents useful forthe suppression of schizophrenia, ADHD and bipolar disorder. The presentinvention further relates to the construction of animal models ofschizophrenia, ADHD and bipolar disorder. Another aspect of theinvention relates generally to neuropsychiatric disorders displaying asimilar dysfunction of the hypothalamo-pituitary-adrenal axis asobserved in the repeated variable prenatal stress animal preparationdescribed herein, such as, schizophrenia, ADHD and bipolar disorder,where normalization of the function of the HPA-axis or acute stressresponse may have therapeutic benefits, or can be used to monitordisease progression and/or treatment response.

BACKGROUND OF THE INVENTION

In many psychiatric disorders, stress is the major non-genetic factorcontributing to the outbreak of the disease, to the manifestation orexacerbation of acute symptoms, to the recurrence or relapse after aperiod of remission and to the failure to respond to pharmacotherapy(e.g. Norman and Malla, Br J Psychiatry. 1993; 162:161-166; Peralta andCuesta, Compr Psychiatry. 1994:198-204; Jansen et al.,Psychopharmacology (Berl). 2000; 149:319-325.)

Schizophrenia is a devastating neuropsychiatric disorder with a 1%worldwide prevalence (Carpenter W T Jr and Buchanan R W, N Engl J Med.330:681-690 (1994); Hyman S E, Biol Psychiatry. 47: 1-7 (2000)). Itsetiology is multifactorial with a complex genetic component and severaldisease loci identified so far (for review, see Pulver, Biol Psychiatry.47:221-30. (2000)). A high discordance among twins, and epidemiologicalstudies suggest the involvement of environmental factors in the originof the disease (Weinberger, Lancet 346: 552-557 (1995); Hyman, BullWorld Health Organ. 78:455-63 (2000), Tsuang et al., Br J Psychiatry(Suppl). 40:18-24 (2001)). One of the most frequently reportedenvironmental risk factors for developing schizophrenia is a chronicmaternal stress in a form or another: Prenatal loss of the father(Huttunen and Niskanen, Arch Gen Psychiatry. 35:429-431. (1978)),influenza (Mednick et al., Arch Gen Psychiatry, 45:189-192 (1988)), war(van Os and Selten, Br J Psychiatry, 172:324-326 (1998)) or naturaldisaster (Selten, Schizophr Res. 35:243-245 (1999); Kinney et al., JAbnorm Psychol., 108:359-362 (1999); Watson et al., Dev Psychopathol.11:457-466 (1999)). On the other hand, chronic maternal stress duringpregnancy may increase the likelihood for preterm birth, and lower birthweight and anoxia which by themselves have been found to be riskfactorsfor schizophrenia (Jones et al., Am. J. Psychiatry 155:355-364 (1998);Hultman et al., BMJ, 318:421-426 (1999)). Gestational stress has alsobeen found to be associated with higher incidence ofattention-deficit-hyperactivity disorder (ADHD; Clements et al., Ga.Educ. Res. 91:1-14 (1992)), bipolar disorder (Brown et al., Am JPsychiatry., 157:190-195 (2000)) and unipolar depression (Watson et al.,Dev Psychopathol., 11:457-466 (1999); Brown et al., Am J Psychiatry.,157:190-195 (2000)). The nature of the behavioral abnormality caused bymaternal stress is probably determined by the time and duration of thestress in relation to the stage of development of particular neuronalsystem. The timing of the stressful event during the second trimester,in contrast to the first or the third, seems to be critical forschizophrenia (Imamura, Acta Psychiatr Scand 100:344-349 (1999); Myhumanet al., Br J Psychiatry 169:637-40 (1996)) and could also account forthe negative outcomes in some studies (Crow, Br. J. Psychiatry164:588-592 (1994); Westergaard et al., Arch Gen Psychiatry 56:993-998(1999)). Assessment of maternal stress as the cause of the behavioralpathology in the epidemiological studies is often confounded by factorssuch as recall bias, concomitant drug intake, poor nutrition, pregnancysymptoms, ill health and/or small size of the study population. Toisolate gestational stress from other possibly contributing factorswell-controlled studies in experimental animals have been undertakenmostly in rats and in non-human primates (for review, see Weinstock,Progr. Neurobiol. 65: 427-451 (2001)). Maternal stress in rats andmonkeys has been shown to result in decreased social interaction of theprenatally stressed offspring (Ward and Stehm, Physiol. Behav.50:601-605 (1991); Clarke and Schneider, Dev. Psychobiol. 26:293-304(1993)) which is in accordance with human findings (Meijer, ActaPsychiatr. Scand. 72:505-511 (1985); Done et al., BMJ 309:699-703(1994)). Furthermore, in adulthood, most prenatally stressed ratpreparations show increased anxiogenic behaviour (Thompson, Science15:698-699 (1957); Wakshlak and Weinstock, Physiol. Behav. 48:289-292(1990); Poltyrev et al., Dev. Psychobiol. 29:453-462 (1996); Vallée etal., J. Neurosci. 17, 2626-2636 (1997)) and depressive behaviour asmeasured by increased immobility time in the forced swim test (Alonso etal., Physiol. Behav. 50:511-517 (1991); Drago et al., EurNeuropsychopharmacol. 9:239-45 (1999) and Weinstock, In: Myslobodsky, Mand Weiner, I. (Eds.) Contemporary Issues in Modeling Psychopthology.Kluwer, Dordrecht, pp. 45-54 (2000)), increased anhedonia (Keshet andWeinstock, Pharmacol Biochem Behav. 50:413-419 (1995)) and alteredsleep-wake cycle (decreased REM-sleep latency, prolongation of the firstREM-sleep episode and decrease in slow-wave-sleep; Rao et al., Prog.Neuropsychopharmacol Biol Psychiatry. 23: 929-939 (1999)). However, noimpairment of prepulse inhibition (PPI), as a measure of abnormality ofsensory gating (inability to filter out the flow of excessive sensoryinformation reaching consciousness) has so far been observed inprenatally stressed animals. On the contrary, Lehmann et al. (BehavBrain Res. 107: 133-44 (2000)) reported an increase in the PPI in ratsthat during their gestation had been exposed to prenatal restraintstress (3×/week). Recently, Koenig et al. (Schizophrenia Res.49(suppl):92 (2001)) have demonstrated that prenatal stress during thesecond week of gestation failed to alter prepulse inhibition responses,while exposure to stress during the third week of gestation(corresponding to the second trimester in humans in terms of braindevelopment, Bayer et al., Neurotoxicology 14:83-144 (1993)) resulted indisruption of sensorimotor gating as measured by PPI. Isolation rearing,which appears to be a social stressor, also leads to abnormal prepulseinhibition (Varty and Geyer, Behav. Neuroscience 112:1-8 (1998)).Cognitive deficits, disrupted hippocampal anatomy and deficits insensorimotor gating, i.e. PPI and P50, are features commonly observed inschizophrenic patients (Flaum et al., J. Psychiatr. Res. 29:261-276(1995); Bilder et al., Schizophr. Res. 17:47-58 (1995); Freedman et al.,Proc. Natl. Acad. Sci. USA 94:587-592 (1997); Clementz et al., Am. J.Psychiatry 155:1691-1694 (1998); Velakoulis et al., Arch. Gen.Psychiatry 56:133-141 (1999); Stefanis et al., Biol. Psychiatry46:697-702 (1999); Gur et al., Arch. Gen. Psychiatry 57:769-775 (2000);Kupferberg and Heckers, Curr. Opin. Neurobiol. 10:205-210 (2000); Walderet al., Biol. Psychiatry 48:1121-1132 (2000)). Lemaire et al. (Proc.Natl. Acad. Sci. USA 97:11032-11037 (2000)) provided evidence showingmaternal stress in rats resulted in the offspring having smallerhippocampi and spatial learning deficits due to impairment in thelearning-associated neurogenesis in the dentate gyrus.

Prenatal stress increases maternal corticotropin and corticosteroneconcentrations. Corticosterone can readily penetrate the fetal brain(Zarrow et al., 1970) and interact with specific glucocorticoidreceptors that are present during the last week of gestation in the rat(Meaney et al., 1985; Cintra et al., 1993). Glucocorticoid receptors arenuclear hormone receptors which function as ligand-activatedtranscription factors directly mediating transactivation of target genesby binding sequence specific recognition elements (glucocorticoidresponse elements; Whitfield et al., 1999). Glucocorticoid receptors arealso known to interact with multiple transcription factors, such asc-jun, nuclear factor-κB, the TFIID complex, STAT5, and co-activatorsknown to modulate the function of these signaling molecules (Jenkins etal., 2001; Yudt and Cidlowski, 2002). Prenatal stress also increasesmaternal and foetal catecholamine release (Morishima et al., 1978;Roehde et al., 1989), maternal oxytocin and opiold peptides of whichβ-endorphin is able to cross the placenta (Sandman and Kastin, 1981;Neumann et al., 1998). Molecular pathways leading from prenatal stressto the neuroendocrinological, behavioral, molecular and neurochemicalchanges in the adult remain poorly understood. In addition to complexglucocorticoid regulated mechanisms, it is likely that the plasticity ofthe developing brain monoaminergic system participates in these changes(Weinstock, 2001; Welberg and Seckl, 2001, Seckl, 2001).

It has been shown repeatedly that exposure to stress in utero reprogramsthe adult hypothalamo-pituitary-adrenal (HPA)-axis resulting in greaterand prolonged elevation of plasma ACTH and/or corticosterone after acutestress (Peters, Pharmacol. Biochem. Behav. 17:721-726 (1982); Henry, J.Neuroendocrinol. 6:341-345 (1994); McCormic, Brain Res Dev Brain Res.84:55-61 (1995); Barbazanges et al., J. Neurosci. 16:3943-3949 (1996;Vallée et al., J. Neurosci. 17, 2626-2636 (1997); Dugovic et al., JNeurosci. 19:8656-64 (1999)) which may be due to a decrease in type Iand type II glucocorticoid receptors in the hippocampus after prenatalstress (Henry et al., J. Neuroendocrinol. 6:341-345 (1994), Koehl etal., J. Neurobiol. 40:302-315 (1999)). The disturbance of the HPA-axisis consistent with schizophrenia (Yeragani, Can. J. Psychiatry35:128-132 (1990); Goldman et al., Am. J. Psychiatry 150:653-655 (1993);Elman et al., Am. J. Psychiatry 155:979-981 (1998); Newcomer et al.,Biol. Psychiatry 29:855-864 (1998)), generalized anxiety disorder anddepression in humans (Arborelius et al., J Endocrinol. 160:1-12 (1999)).Schizophrenic patients have been also shown to have fewer glucocorticoidreceptors in postmortem schizophrenic brain tissue (Knable et al.,Schizophrenia Res. 49(Suppl):53 (2001)).

Gestational stress has been shown to cause changes in brain morphologysuch as a reduction in hippocampal synapses (Hayashi et al., Int J DevNeurosci. 16: 209-216 (1998)) which may be partially due to the reducedneuronal number in hippocampus as a result of maternal stress as shownby Lemaire et al. (Proc. Natl. Acad. Sci. USA 97:11032-11037 (2000)).Also, lower levels of N-acetyl aspartate (NAA) which is used as a markerfor neuronal metabolism have been reported in the left prefrontal cortex(PFC) of prenatally stressed rats (Poland et al., J Psychiatr Res. 33:41-51 (1999)) which has also be shown for untreated schizophrenic PFCand hippocampus (Buckley et al., Biol Psychiatry. 36:792-800 (1994);Bertolino et al., Biol Psychiatry. 43:641-8 (1998)).

Glucocorticoid hormones or stress have been shown to influenceneurotransmission at many levels. For the brain dopaminergic system,these effects appear to be mediated by glucocorticoid receptors that areexpressed in many dopamine (DA)-containing neurons in the brain(Harfstrand et al., Proc. Natl. Acad. Sci. USA 83:9779-9783 (1986);Cintra et al., Neuroendocrinology 57:1133-1147 (1993)). Glucocorticoidsenhance DA metabolism (Tanganelli et al., J. Neural Transm. Gen. Sect.81:183-194 (1990); Takahashi et al., Brain Res. 574:131-137 (1992); Diazet al., Neuroscience 81:129-140 (1995); Barrot et al., Eur. J. Neurosci.12:973-979 (2000), Eur. J. Neurosci. 13:812-818 (2001)) in the striatumand increase DA release in the nucleus accumbens (Piazza et al., Proc.Natl. Acad. Sci. USA 93:8716-8720 (1996); Barrot et al., Eur. J.Neurosci. 12:973-979 (2000)), although also decreased DA metabolism inthe nucleus accumbens has been reported (Alonso et al., Pharmacol.Biochem. Behav. 49:353-358 (1994)). Furthermore, increased D2 receptorbinding has been reported in the nucleus accumbens (Henry et al., BrainRes. 685:179-186 (1995)). In human subjects, glucocorticoidadministration increases plasma HVA levels while inducing psychoticsymptoms (Wolkowitz, Psychoneuroendocrinology 19:233-255 (1994)). Inschizophrenic patients, augmentation in the dopaminergic state has beenbeautifully demonstrated by Laruelle et al. (Proc. Natl. Acad. Sci. USA93:9235-9240 (1996), Biol. Psychiatry 46:56-72 (1999)).

In addition to changes in the dopaminergic system, exposure toglucocorticoids causes an increase in the turnover of norepinephrine inthe cortex and locus coeruleus (Takahashi et al., Brain Res. 574:131-137(1992)). It also increases serotonin synthesis in fetal brain (Peters,Pharmacol. Biochem. Behav. 25:873-877 (1986), Pharmacol. Biochem. Behav.31:839-843 (1988), Pharmacol. Biochem. Behav. 35:943-947 (1990)), aswell as concentrations of the brainstem serotonin transporter in rats(Slotkin et al., Brain Res. Dev. Brain Res. 93:155-161 (1996)). Duringrat brain development, glucocorticoid receptors first appear onembryonic day (E) 13 (Cintra et al., Neuroendocrinology 57:1133-1147(1993)) which coincides the developmental phase of all the monoaminergicneurotransmitter systems. Therefore, early exposure to glucocorticoidsalso modifies the development of neurotransmitter systems proposed to beinvolved in the pathophysiology of schizophrenia, includingnorepinephrine, serotonin, dopamine and GABA (Davis et al., Am JPsychiatry 148:1474-1486 (1991); Roth and Meltzer, In Bloom F E andKupfer D J (Eds), Psychopharmacology: The fourth generation of progress,New York, Raven Press, pp 1215-1227 (1995); Muneoka et al., Am. J.Physiol. 273:R1669-1675 (1997); Slotkin et al., Brain Res. Dev. BrainRes. 93:155-161 (1996); Diaz et al., Neuroscience 66:467-473. (1995);Neuroscience 81:129-140 (1997); Lewis, Brain Res Brain Res Rev.31:270-6. (2000).

While effects of prenatal stress on the HPA axis and on some of theneurotransmitter systems are relatively well known, and the concept ofprenatal programming has been a long-standing topic inneuroendocrinology research (for reviews, see e.g. Weinstock, Progr.Neurobiol. (2001); Weldberg and Seckl, J. Neuroendocrinolol., 13:113-128(2001)) the molecular mechanisms resulting in and from the reprogrammingremain poorly understood and, changes at the global gene expressionlevel have not been studied previously. The hypothesis (see Koenig etal., Neuropsychopharmacol. 27: 309-318 (2002)) that prenatalhypercortisolemia coinciding a critical neurodevelopmental period in theembryo increases the risk of schizophrenia and other disorders withneurodevelopmental component (such as bipolar disorder and ADHD) hasbeen shown to be biologically plausible as analogous changes can beobserved in animals under controlled circumstances. Prenatal stressprovides a testable preclinical and clinical model for underlyingmolecular pathophysiology and cure of the above mentioned humandisorders.

Antipsychotic drugs were identified in the 1950's, and these drugs werefound to produce a dramatic improvement in the psychotic phase of theillness. Reserpine was the first of these drugs to be used and wasfollowed by typical antipsychotic drugs including phenothiazines, thebutyrophenones, and the thioxanthenes. A new group of therapeutic drugs,typified by clozapine, has been developed and were referred to as“atypical” antipsychotics. Haloperidol has been employed extensively inthe treatment of schizophrenia and is one of the currently preferredoptions for treatment. When these drugs are taken over the course of atleast several weeks, they mitigate or eliminate delusions,hallucinations, and some types of disordered thinking. Maintenance of apatient on these drugs reduces the rate of relapse. Since there is noway of determining if an individual is susceptible to schizophrenia, itis currently unknown if these antipsychotic compounds are useful in theprophylactic treatment of schizophrenia.

Signal transduction is the general process by which cells respond toextracellular signals (e.g. neurotransmitters) through a cascade ofbiochemical reactions. The first step in this process is the binding ofa signaling molecule to a cell membrane receptor that typically leads tothe inhibition or activation of an intracellular enzyme. This type ofprocess regulates many cell functions including cell proliferation,differentiation, and gene transcription (WO 01/26622). One importantmechanism by which signal transduction occurs is through G-proteins.Receptors on the cell surface are coupled intracellularly to a G-proteinthat becomes activated, when the receptor is occupied by an agonist, bybinding to the molecule GTP. G-proteins may influence a large number ofprocesses including voltage-activated calcium channels, adenylatecyclase, and phospholipase C.

There is a need in the art for the identification of schizophrenia, ADHDand bipolar disorder associated genes. Identification of such geneswould provide a fundamental understanding of the disease process fromwhich a number of clinically important applications would arise. Saidgenes identified may lead to the development of therapeutics (smallmolecule drugs, antisense molecules, antibody molecules) directlytargeted to the gene or protein product of the gene, or may target thebiochemical pathway of which the protein product is a part at anupstream or downstream location if the development of such drugs iseasier than directly targeting the gene or its protein product.Polynucleotide sequences comprising the gene, sequence variants thereofand protein products thereof may be used to develop a clinicaldiagnostic test for schizophrenia, ADHD and bipolar disorder and for theidentification of individuals at high risk for the development ofschizophrenia, ADHD and bipolar disorder. The results of such tests mayalso have prognostic value and may be used to predict patients whorespond to and those who do not respond to drug therapy.

Prefrontal cortex, which is part of the frontal pole, is a target forglucocorticoids involved in stress response (Meaney M J and Aitken D H,Brain Res. 328:176-180. (1985)), it regulates the negativeglucocorticoid feedback of the HPA-axis (Diorio D. et al., J. Neurosci.13:3839-3847 (1993); Moghaddam B., Biol Psychiatry. 51, 775-787.(2001)), it shows neurochemical and neuroanatomical changes in responseto stress (Wellman C., J. Neurobiol. 49:245-253 (2001)), and mediatesmany of the behaviors that are altered by chronic stress orcorticosterone treatment, for example memory and learning.Identification of the abnormal stress response pattern at geneexpression level in the frontal pole is relevant with regard to anystress provoked or stress-deteriorating human condition where frontalcortical structures play a role as parts of circuitry, such asdepression, bipolar disorder, schizophrenia, anxiety, panic disorder,generalized anxiety disorder, post-traumatic stress syndrome, bipolardisorder, and addiction, and has got a potential for identification ofnovel therapeutic approaches that aim at normalizing the abnormal stressresponse in order to alleviate the resulting conditions, or asbiomarkers for therapeutic interventions that also normalize theabnormal stress response.

SUMMARY OF THE INVENTION

In a first aspect, the invention provides the use of isolated DNAs inthe prevention and treatment of schizophrenia, ADHD and/or bipolardisorder comprising the nucleotide sequences referenced in Tables 1 and2. Also provided are the use of isolated DNA's that comprise nucleicacid sequences in said indications that hybridize under high stringencyconditions to the isolated DNA referenced in Tables 1 and 2 (hereinreferred to as the “Genes of the invention”). Also provided are the useof said nucleic acid sequences comprising at least about 15 bases,preferably at least about 20 bases, more preferably a nucleic acidsequence comprising about 30 contiguous bases the nucleic acid sequencereferenced in Tables 1 and 2.

In a second aspect, the invention provides the use of isolatedpolypeptides in the prevention and treatment of schizophrenia, ADHDand/or bipolar disorder with the amino acid sequences encoded by theGenes of the invention. Such polypeptides, or fragments thereof, arefound e.g. in the prefrontal pole and/or hypothalamus of sufferers ofschizophrenia, ADHD and/or bipolar disorder in a different amount thanin the same tissues of individuals without schizophrenia, ADHD and/orbipolar disorder.

A third aspect of the present invention encompasses a method for thediagnosis of schizophrenia, ADHD and/or bipolar disorder in a humanwhich includes detecting the elevated and/or lowered transcription ofmessenger RNA transcribed from the Genes of the invention, where suchaltered transcription is diagnostic of the human's affliction withschizophrenia, ADHD and/or bipolar disorder. Another embodiment of theassay aspect of the invention provides a method for the diagnosis ofschizophrenia, ADHD and/or bipolar disorder in a human which requiresmeasuring the amount of a polypeptide that include or are thepolypeptides which are encoded by the Genes of the invention, e.g. inthe frontal pole and/or hypothalamus from a human, where the presence ofan elevated or lowered amount of the polypeptide or fragments thereof,relative to the amount of the polypeptide or fragments thereof in e.g.in frontal pole and/or hypothalamus from a healthy human, is diagnosticof the human's suffering from schizophrenia, ADHD and/or bipolardisorder. A yet further aspect of the present invention is a method ofdetermining susceptibility to schizophrenia, ADHD and/or bipolardisorder comprising obtaining from a patient to be tested forsusceptibility to schizophrenia, ADHD and/or bipolar disorder a sampleof tissue, measuring levels of the polypeptides encoded by the Genes ofthe invention in said sample, and determining if there is an elevated orlowered level of the polypeptides encoded by the Genes of the inventionin the sample. Another aspect of the present invention is a kit fordiagnosing schizophrenia, ADHD and/or bipolar disorder in a patient,said kit comprising antibodies to the polypeptides encoded by the Genesof the invention, and a detector for ascertaining whether saidantibodies bind to the polypeptides encoded by the Genes of theinvention in a sample. Another aspect of the present invention is a kitfor diagnosing schizophrenia, ADHD and/or bipolar disorder in a patient,said kit comprising a detection tool to measure transcript levels of theGenes of the invention in a patient, and of a standard to ascertainaltered levels of transcript of the Genes of the invention in thepatient.

In another aspect, the invention is directed to methods for theidentification of molecules that can bind to the polypeptides encoded bythe Genes of the invention and/or modulate, i.e. activate or inhibit theactivity of the polypeptides encoded by the Genes of the invention ormolecules that can bind to nucleic acid sequences of the Genes of theinvention that modulate the transcription or translation of said Genes.Such methods are disclosed in, e.g., U.S. Pat. Nos. 5,541,070;5,567,317; 5,593,853; 5,670,326; 5,679,582; 5,856,083; 5,858,657;5,866,341; 5,876,946; 5,989,814; 6,010,861; 6,020,141; 6,030,779; and6,043,024, all of which are incorporated by reference herein in theirentirety. Molecules identified by such methods also fall within thescope of the present invention.

Yet another aspect of the present invention is a method of treatingschizophrenia, ADHD and/or bipolar disorder said method comprisingmeasuring levels of the Polypeptides of the invention or levels of mRNAof the Genes of invention in a patient, and altering said polypeptidelevels to provide the patient with an improved psychiatric function. Inaccordance with one embodiment of this aspect of the invention there areprovided antisense polynucleotides that regulate transcription of theGenes of the invention; in another embodiment, double stranded RNA isprovided that can regulate the transcription of the Genes of theinvention.

An additional aspect of the present invention is an animal model forschizophrenia, ADHD and/or bipolar disorder where a repeated variablematernal stress was applied on Sprague Dawley rats during thegestational days 14-22, coinciding with the second trimester in humans,as opposed to repeated administration of the same stress to the pregnantanimal.

A still additional aspect of the invention is the use of the genesidentified in Tables 5, 6 and 7 in treating schizophrenia, ADHD and/orbipolar disorder and/or other stress aggravated psychiatric conditionwhere dysregulation of the normal HPA axis function is a clinicalfinding (dysregulation of the HPA axis in this instance is defined asprolongation of the corticosterone response after acute stress or asinsensitivity to glucocorticoid negative feedback. In humans, thiscorresponds to an “escape” from dexamethasone induced suppression of theHPA axis or abnormal elevation of diurnal cortisol in the a.m. or p.m.in patients); methods comprising measuring levels of the Polypeptides ofthe invention or levels of mRNA of the Genes of invention in a patient,and altering said polypeptide levels to provide the patient with animproved psychiatric function. Another dimension is the use of genesidentified in Tables 5-7 as biomarkers of the disease progression ortreatment response.

Other objects, features, advantages and aspects of the present inventionwill become apparent to those of skill from the following description.It should be understood, however, that the following description and thespecific examples, while indicating preferred embodiments of theinvention, are given by way of illustration only. Various changes andmodifications within the spirit and scope of the disclosed inventionwill become readily apparent to those skilled in the art from readingthe following description and from reading the other parts of thepresent disclosure.

DESCRIPTION OF THE INVENTION

All patent applications, patents and literature references cited hereinare hereby incorporated by reference in their entirety.

In practicing the present invention, many conventional techniques inmolecular biology, microbiology, and recombinant DNA are used. Thesetechniques are well known and are explained in, for example, CurrentProtocols in Molecular Biology, Volumes I, II, and III, 1997 (F. M.Ausubel ed.); Sambrook et al., 1989, Molecular Cloning: A LaboratoryManual, Second Edition, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y.; DNA Cloning: A Practical Approach, Volumes I and II, 1985(D. N. Glover ed.); Oligonucleotide Synthesis, 1984 (M. L. Gait ed.);Nucleic Acid Hybridization, 1985, (Hames and Higgins); Transcription andTranslation, 1984 (Hames and Higgins eds.); Animal Cell Culture, 1986(R. I. Freshney ed.); Immobilized Cells and Enzymes, 1986 (IRL Press);Perbal, 1984, A Practical Guide to Molecular Cloning; the series,Methods in Enzymology (Academic Press, Inc.); Gene Transfer Vectors forMammalian Cells, 1987 (J. H. Miller and M. P. Calos eds., Cold SpringHarbor Laboratory); and Methods in Enzymology Vol. 154 and Vol. 155 (Wuand Grossman, and Wu, eds., respectively).

The present invention relates to the identification of genes that areexpressed at higher or lower levels in the frontal lobe and/orhypothalamus in a patient with schizophrenia, ADHD and/or bipolardisorder, than as in the same tissue in a healthy individual. By virtueof their low or high expression in said normal body tissues includingnormal frontal lobe and/or hypothalmic tissues and their high or lowexpression in said tissues in a patient with schizophrenia, ADHD and/orbipolar disorder these genes can be utilized in the diagnosis, diseasemanagement, screening for agonists and antagonists of the Genes of theinvention, disease prevention and treatment, and/or post-treatmentfollow-up of men at risk for, with, or at risk for recurrence of saiddisorders which include, but are not limited to, schizophrenia, ADHDand/or bipolar disorder. These Genes of the invention include thoselisted in Tables 1 and 2 and are for the first time, as to ourknowledge, associated with schizophrenia, ADHD and/or bipolar disorderand those listed in Tables 5, 6 and 7 for the first time associated witheither normal or abnormal stress response related to disruption ofHPA-axis. The complete sequences of the all the Genes of the inventionare available from public databases (NCBI) using the accession numbersshown in Table 1, 2, 5, 6, and 7.

Although stress is commonly used to exacerbate behavioral abnormalitiesin psychiatric animal models (e.g. Piazza and Le Moal, Trends PharmacolSci. 1998; 19:67-74) abnormal stress responses in general, have not beenincorporated in the animal models themselves. In an attempt to searchfor a novel heuristic animal model for schizophrenia we have incorporatestress in a form of repeated variable prenatal stress to pregnantSprague-Dawley rats in a randomized manner during the last week ofpregnancy (Koenig et al., 2002, Society for Neuroscience Abstracts,495.6; Kinnunen et al., 2002, Society for Neuroscience Abstracts,495.7). This in terms of brain development, corresponds to the secondtrimester in humans (Bayer et al., 1993). The offspring is leftundisturbed until weaning and behavioral testing on days 35 and 56. Thelocomotor response to amphetamine and phencyclidine (PCP) is enhanced inthe prenatally stressed offspring on postnatal day 56 but not onpostnatal day 35. The prenatally stressed male offspring displayed adisruption of sensorimotor gating as measured by prepulse inhibition(PPI) and in sensory gating assessed using the N40 response and adisruption in the normal corticosteroid negative feedback are commonlyobserved deficits in schizophrenic patients. Also the emergence of thebehavioral deficits postpubertally is in accordance with schizophrenia.

Using said microarray analysis approach genes were identified that inthe frontal pole are differentially regulated following an acute stressexposure. Based on the literature, some of these genes are known to beregulated by acute stress or corticosterone, e.g. c-fos (e.g. Martinezet al., Stress 2002; 5:3-13), sgk (Webster et al., Mol. Cell. Biol.,1993; 13:2031-2040; Naray-Fejes-Toth et al., J. Steroid Biochem. Mol.Biol., 2000, 75:51-56) and CART (Vrang et al., Brain Res., 2003, 965:45-50). The gene expression response that is occurring in the normalcontrol animals and is partially overlapping with the acute stressresponse in the prenatally stressed animals, provides a pattern for“normal stress response” in adult Sprague-Dawley male rats. Furthermore,by the present invention genes were identified that are differentiallyregulated by acute stress in the prenatally stressed animals only, orare differentially regulated by acute stress following a different timecourse than in the normal controls. Those genes that are differentiallyregulated after the acute stress in the prenatally stressed thereforedemarcates the gene expression component of “abnormal stress response”in adult Sprague-Dawley male rats.

Any selection of at least one of the genes listed in Tables 1, 2, and/or3 can be utilized as a marker and/or therapeutic target forSchizophrenia, bipolar disorder, and/or ADHD with the proviso that ifexpression of only one gene is detected that the gene is not one of thegenes of Table 3. In some embodiments, if expression of only one gene isdetected the gene is not one of Table 3 which are already known to belinked with schizophrenia, bipolar disorder, ADHD. In preferredembodiments, any selection of at least one of the Genes of the inventioncan be utilized as a marker and/or therapeutic target for schizophrenia,bipolar disorder, ADHD. In particularly useful embodiments, the genes inTables 1 and/or 2 can be utilized as a therapeutic target forschizophrenia, bipolar disorder, ADHD. In particularly usefulembodiments, a plurality of these genes or at least two or more of thegenes listed in Table 1, 2, and/or 3 and Table 5, 6 and/or 7 can beselected and their expression monitored simultaneously to provideexpression profiles for use in various aspects. For example, expressionprofiles of the genes provide valuable molecular tools for rapidlydiagnosing and monitoring the progression of schizophrenia, bipolardisorder, and/or ADHD, and for evaluating drug efficacy. Changes in theexpression profile from a baseline profile can be used as an indicationof such effects. Accordingly, the invention provides methods forscreening a subject for (diagnostic) schizophrenia, bipolar disorder,and/or ADHD or at risk of developing (prognostic) schizophrenia, bipolardisorder, and/or ADHD, methods for monitoring the progression ofschizophrenia, bipolar disorder, and/or ADHD in a subject, methods forthe identification of agents that are useful in treating a subjecthaving or at risk of having schizophrenia, bipolar disorder, and/orADHD, methods of treating a subject having or at risk of havingschizophrenia, bipolar disorder, and/or ADHD, methods for monitoring theefficacy of certain drug treatments for schizophrenia, bipolar disorder,and/or ADHD, and vectors for schizophrenia, bipolar disorder, and/orADHD-specific replication.

The most differentially expressed genes are identified in Tables 1, 2and/or 3. Particularly preferred genes listed in Table 1 and/or 2include genes involved with synaptic vesicle endo- and exocytosis, geneswhich are found to be in the postsynaptic density, some of the NMDA,AMPA- and GABA receptor subunits.

The method for screening a subject for schizophrenia, bipolar disorder,and/or ADHD or at risk of developing schizophrenia, bipolar disorder,and/or ADHD comprises:

-   -   a) detecting a level of expression of at least one of the genes        identified in Tables 1, 2, and/or 3 in a sample of brain tissue,        e.g. frontal pole and/or hypothalamus obtained from a mammal,        e.g. human or animal model to provide a first value; and    -   b) comparing the first value with a level of expression of the        at least one gene identified in Tables 1, 2, and/or 3 in a        sample of fluid or tissue as above obtained from a subject free        of schizophrenia, bipolar disorder, and/or ADHD, wherein a        greater or lesser, expression level in the subject sample,        compared to the sample from the subject free of schizophrenia,        bipolar disorder, and/or ADHD, is indicative of the subject        having schizophrenia, bipolar disorder, and/or ADHD or at risk        of developing schizophrenia, bipolar disorder, and/or ADHD.

The brain tissue sample (e.g. from the frontal pole and/or hypothalamus)can be obtained from the subject, a human or animal model, by knownsurgical methods, e.g., surgical resection or needle biopsy. Whenassessing the level of expression of the gene by measuring the level ofmRNA as described below, it is preferable to obtain a brain tissuesample, e.g. frontal pole and/or hypothalamus, from the subject. Thesample taken from the subject free from schizophrenia, bipolar disorder,and/or ADHD can be a sample of normal brain tissue (e.g. frontal poleand/or hypothalamus) or bodily fluid from the same individual or fromanother individual. For example, in examination of a suspected disordersuch as schizophrenia, bipolar disorder, and/or ADHD, the sample fromthe subject free from schizophrenia, bipolar disorder, and/or ADHD(disease-free subject) can be a sample of normal brain cells from theindividual suspected of having schizophrenia, bipolar disorder, and/orADHD. The sample obtained from the disease-free subject can be obtainedat the same time as the sample obtained from the subject, or can be apre-established control for which expression of the gene was determinedat an earlier time. The level of expression of the gene in the sampleobtained from the disease-free subject is determined and quantitatedusing the same approach as used for the sample obtained from thesubject. The level of expression of at least one of the disclosed genesin the samples obtained from the subject and disease-free subject can bedetected by measuring either the level of mRNA corresponding to thegene, the protein encoded by the gene or a fragment of the protein,e.g., the catalytic domain. In the methods of the invention, the levelof expression of one of the disclosed genes in a diseased tissuepreferably differs from the level of expression of the gene in anon-diseased tissue by a statistically significant amount. In presentlypreferred embodiments, at least about a 1.5-fold difference inexpression levels is observed. In some embodiments, the expressionlevels of a gene differ by at least about 2-, 3-, 4-, 5-, 10- or100-fold or more in the diseased tissue compared to the non-diseasedtissue. The level of expression of at least one of the genes disclosedin Table 1 is determined in the methods of the invention. It issometimes desirable to determine the level of expression of 2, 3, 5, 10,20, or more of the disclosed genes. RNA can be isolated from the samplesby methods well known to those skilled in the art as described, e.g., inAusubel et al., Current Protocols in Molecular Biology, Vol. 1, pp.4.1.1-4.2.9 and 4.5.1-4.5.3, John Wiley & Sons, Inc. (1996). Methods fordetecting the level of expression of mRNA are well known in the art andinclude, but are not limited to, northern blotting, reversetranscription PCR, real time quantitative PCR and other hybridizationmethods. A particularly useful method for detecting the level of mRNAtranscripts obtained from a plurality of the disclosed genes involveshybridization of labeled mRNA to an ordered array of oligonucleotides.Such a method allows the level of transcription of a plurality of thesegenes to be determined simultaneously to generate gene expressionprofiles or patterns. The gene expression profile derived from thesample obtained from the subject can be compared with the geneexpression profile derived from the sample obtained from thedisease-free subject to determine whether the genes are over-expressedin the sample from the subject relative to the genes in the sampleobtained from the disease-free subject, and thereby determine whetherthe subject has or is at risk of developing schizophrenia, bipolardisorder and ADHD. The oligonucleotides utilized in this hybridizationmethod typically are bound to a solid support. Examples of solidsupports include, but are not limited to, membranes, filters, slides,paper, nylon, wafers, fibers, magnetic or nonmagnetic beads, gels,tubing, polymers, polyvinyl chloride dishes, etc. Any solid surface towhich the oligonucleotides can be bound, either directly or indirectly,either covalently or non-covalently, can be used. A particularlypreferred solid substrate is a high density array or DNA chip (seeExample 2). These high density arrays contain a particularoligonucleotide probe in a pre-selected location on the array. Eachpre-selected location can contain more than one molecule of theparticular probe. Because the oligonucleotides are at specifiedlocations on the substrate, the hybridization patterns and intensities(which together result in a unique expression profile or pattern) can beinterpreted in terms of expression levels of particular genes. Theoligonucleotide probes are preferably of sufficient length tospecifically hybridize only to complementary transcripts of the aboveidentified gene(s) of interest. As used herein, the term“oligonucleotide” refers to a single-stranded nucleic acid. Generallythe oligonucleotides probes will be at least 16 to 20 nucleotides inlength, although in some cases longer probes of at least 20 to 25nucleotides will be desirable. The oligonucleotide probes can be labeledwith one or more labeling moieties to permit detection of the hybridizedprobe/target polynucleotide complexes. Labeling moieties can includecompositions that can be detected by spectroscopic, biochemical,photochemical, bioelectronic, immunochemical, electrical optical orchemical means. Examples of labeling moieties include, but are notlimited to, radioisotopes, e.g., ³²P, ³³P, ³⁵S, chemiluminescentcompounds, labeled binding proteins, heavy metal atoms, spectroscopicmarkers such as fluorescent markers and dyes, linked enzymes, massspectrometry tags, and magnetic labels. Oligonucleotide probe arrays forexpression monitoring can be prepared and used according to techniqueswhich are well known to those skilled in the art as described, e.g., inLockhart et al., Nature Biotechnology, Vol. 14, pp. 1675-1680 (1996);McGall et al., Proc. Natl. Acad. Sci. USA, Vol. 93, pp. 13555-13460(1996); and U.S. Pat. No. 6,040,138.

Expression of the protein encoded by the gene(s) or a fragment of theprotein, e.g., the catalytic domain, can be detected by a probe which isdetectably labeled, or which can be subsequently labeled. Generally, theprobe is an antibody which recognizes the expressed protein. As usedherein, the term antibody includes, but is not limited to, polyclonalantibodies, monoclonal antibodies, humanized or chimeric antibodies andbiologically functional antibody fragments, which are those fragmentssufficient for binding of the antibody fragment to the protein or afragment of the protein. For the production of antibodies to a proteinencoded by one of the disclosed genes or to a fragment of the protein,various host animals may be immunized by injection with the polypeptide,or a portion thereof. Such host animals may include, but are not limitedto, rabbits, mice and rats, to name but a few. Various adjuvants may beused to increase the immunological response, depending on the hostspecies, including, but not limited to, Freund's (complete andincomplete), mineral gels such as aluminum hydroxide, surface activesubstances such as lysolecithin, pluronic polyols, polyanions, peptides,oil emulsions, keyhole limpet hemocyanin, dinitrophenol, and potentiallyuseful human adjuvants such as BCG (bacille Calmette-Guerin) andCorynebacterium parvum. Polyclonal antibodies are heterogeneouspopulations of antibody molecules derived from the sera of animalsimmunized with an antigen, such as target gene product, or an antigenicfunctional derivative thereof. For the production of polyclonalantibodies, host animals, such as those described above, may beimmunized by injection with the encoded protein, or a portion thereof,supplemented with adjuvants as also described above. Monoclonalantibodies (mAbs), which are homogeneous populations of antibodies to aparticular antigen, may be obtained by any technique which provides forthe production of antibody molecules by continuous cell lines inculture. These include, but are not limited to, the hybridoma techniqueof Kohler and Milstein (Nature, Vol. 256, pp. 495-497 (1975); and U.S.Pat. No. 4,376,110), the human B-cell hybridoma technique (Kosbor etal., Immunology Today, Vol. 4, p. 72 (1983); Cole et al., Proc. Natl.Acad. Sci. USA, Vol. 80, pp. 2026-2030 (1983)), and the EBV-hybridomatechnique (Cole et al., Monoclonal Antibodies and Cancer Therapy, AlanR. Uss, Inc., pp. 77-96 (1985)). Such antibodies may be of anyimmunoglobulin class, including IgG, IgM, IgE, IgA, IgD, and anysubclass thereof. The hybridoma producing the mAb of this invention maybe cultivated in vitro or in vivo. Production of high titers of mAbs invivo makes this the presently preferred method of production. Inaddition, techniques developed for the production of “chimericantibodies” (Morrison et al., Proc. Natl. Acad. Sci. USA, Vol. 81, pp.6851-6855 (1984); Neuberger et al., Nature, Vol. 312, pp. 604-608(1984); Takeda et al., Nature, Vol. 314, pp. 452-454 (1985)) by splicingthe genes from a mouse antibody molecule of appropriate antigenspecificity, together with genes from a human antibody molecule ofappropriate biological activity, can be used. A chimeric antibody is amolecule in which different portions are derived from different animalspecies, such as those having a variable or hypervariable region derivedfrom a murine mAb and a human immunoglobulin constant region.Alternatively, techniques described for the production of single-chainantibodies (U.S. Pat. No. 4,946,778; Bird, Science, Vol. 242, pp.423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA, Vol. 85, pp.5879-5883 (1988); and Ward et al., Nature, Vol. 334, pp. 544-546 (1989))can be adapted to produce differentially expressed gene-single chainantibodies. Single chain antibodies are formed by linking the heavy andlight chain fragments of the Fv region via an amino acid bridge,resulting in a single-chain polypeptide. Most preferably, techniquesuseful for the production of “humanized antibodies” can be adapted toproduce antibodies to the proteins, fragments or derivatives thereof.Such techniques are disclosed in U.S. Pat. Nos. 5,932,448; 5,693,762;5,693,761; 5,585,089; 5,530,101; 5,569,825; 5,625,126; 5,633,425;5,789,650; 5,661,016; and 5,770,429. Antibody fragments which recognizespecific epitopes may be generated by known techniques. For example,such fragments include, but are not limited to, the F(ab′)₂ fragments,which can be produced by pepsin digestion of the antibody molecule, andthe Fab fragments, which can be generated by reducing the disulfidebridges of the F(ab′)₂ fragments. Alternatively, Fab expressionlibraries may be constructed (Huse et al., Science, Vol. 246, pp.1275-1281 (1989)) to allow rapid and easy identification of monoclonalFab fragments with the desired specificity. The extent to which theknown proteins are expressed in the sample is then determined byimmunoassay methods which utilize the antibodies described above. Suchimmunoassay methods include, but are not limited to, dot blotting,western blotting, competitive and noncompetitive protein binding assays,enzyme-linked immunosorbant assays (ELISA), immunohistochemistry,fluorescence-activated cell sorting (FACS), and others commonly used andwidely described in scientific and patent literature, and many employedcommercially. Particularly preferred, for ease of detection, is thesandwich ELISA, of which a number of variations exist, all of which areintended to be encompassed by the present invention. For example, in atypical forward assay, unlabeled antibody is immobilized on a solidsubstrate and the sample to be tested is brought into contact with thebound molecule and incubated for a period of time sufficient to allowformation of an antibody-antigen binary complex. At this point, a secondantibody, labeled with a reporter molecule capable of inducing adetectable signal, is then added and incubated, allowing time sufficientfor the formation of a ternary complex of antibody-antigen-labeledantibody. Any unreacted material is washed away, and the presence of theantigen is determined by observation of a signal, or may be quantitatedby comparing with a control sample containing known amounts of antigen.Variations on the forward assay include the simultaneous assay, in whichboth sample and antibody are added simultaneously to the bound antibody,or a reverse assay, in which the labeled antibody and sample to betested are first combined, incubated and added to the unlabeled surfacebound antibody. These techniques are well known to those skilled in theart, and the possibility of minor variations will be readily apparent.As used herein, “sandwich assay” is intended to encompass all variationson the basic two-site technique. For the immunoassays of the presentinvention, the only limiting factor is that the labeled antibody be anantibody which is specific for the protein expressed by the gene ofinterest, or a fragment thereof. The most commonly used reportermolecules in this type of assay are either enzymes, fluorophore- orradionuclide-containing molecules. In the case of an enzyme immunoassay,an enzyme is conjugated to the second antibody, usually by means ofglutaraldehyde or periodate. As will be readily recognized, however, awide variety of different ligation techniques exist which are well-knownto the skilled artisan. Commonly used enzymes include horseradishperoxidase, glucose oxidase, beta-galactosidase and alkalinephosphatase, among others. The substrates to be used with the specificenzymes are generally chosen for the production, upon hydrolysis by thecorresponding enzyme, of a detectable color change. For example,p-nitrophenyl phosphate is suitable for use with alkaline phosphataseconjugates; for peroxidase conjugates, 1,2-phenylenediamine or toluidineare commonly used. It is also possible to employ fluorogenic substrates,which yield a fluorescent product, rather than the chromogenicsubstrates noted above. A solution containing the appropriate substrateis then added to the tertiary complex. The substrate reacts with theenzyme linked to the second antibody, giving a qualitative visualsignal, which may be further quantitated, usuallyspectrophotometrically, to give an evaluation of the amount of secretedprotein or fragment thereof. Alternately, fluorescent compounds, such asfluorescein and rhodamine, may be chemically coupled to antibodieswithout altering their binding capacity. When activated by illuminationwith light of a particular wavelength, the fluorochrome-labeled antibodyabsorbs the light energy, inducing a state of excitability in themolecule, followed by emission of the light at a characteristic longerwavelength. The emission appears as a characteristic color visuallydetectable with a light microscope. Immunofluorescence and EIAtechniques are both very well established in the art and areparticularly preferred for the present method. However, other reportermolecules, such as radioisotopes, chemiluminescent or bioluminescentmolecules may also be employed. It will be readily apparent to theskilled artisan how to vary the procedure to suit the required use.

In another aspect, kits are provided for detecting the level ofexpression of at least one gene identified in Tables 1, 2, and/or 3 in abiological sample, e.g., brain tissue (e.g. frontal pole and/orhypothalamus), with the proviso that if expression of only one gene isdetected that the gene is not any one of the genes of Table 3. In someembodiments, if expression of only one gene is detected the gene is notany one of the genes of Table 3. For example, the kit can comprise alabeled compound or agent capable of detecting a protein encoded by, ormRNA corresponding to, at least one of the genes identified in Tables 1,2, and/or 3 or fragment of the protein, means for determining the amountof protein encoded by or mRNA corresponding to the gene or fragment ofthe protein; and means for comparing the amount of protein encoded by ormRNA corresponding to the gene or fragment of the protein, obtained fromthe subject sample with a standard level of expression of the gene,e.g., from a disease-free subject. The compound or agent can be packagedin a suitable container. The kit can further comprise instructions forusing the kit to detect protein encoded by or mRNA corresponding to thegene.

In another aspect, progression of schizophrenia, bipolar disorder and/orADHD in a subject can be monitored by measuring a level of expression ofmRNA corresponding to, or protein encoded by, at least one of the genesidentified in Tables 1, 2, and/or 3 in a sample of brain tissue (e.g.frontal pole and/or hypothalamus) obtained in the subject over time,i.e., at various stages of the disorder, with the proviso that ifexpression of only one gene is detected that the gene is not any one ofthe genes of Table 3, measuring level of expression of mRNAcorresponding to, or protein encoded by, at least one of the genesidentified in Tables 5, 6 and/or 7 after an exposure to acute stress ore.g. dexamethasone. In some embodiments, if expression of only one geneis detected the gene is not any one of the genes of Table 3. Analteration, i.e. increase or decrease (depending on the diseaseassociated up- or down-regulation of the Genes of the invention) in thelevel of expression of the mRNA or encoded protein corresponding to thegene(s) over time is indicative of the progression of schizophrenia,bipolar disorder and/or ADHD. The level of expression of mRNA andprotein corresponding to the gene(s) can be detected by standard methodsas described above. In a particularly useful embodiment, the level ofmRNA expression of a plurality of the disclosed genes, at least two ofthe Genes of the invention, can be measured simultaneously in a subjectat various stages of schizophrenia, bipolar disorder and/or ADHD togenerate a transcriptional or expression profile of schizophrenia,bipolar disorder and/or ADHD over time. For example, mRNA transcriptscorresponding to a plurality of these genes can be obtained from braincells (e.g. from frontal pole and/or hypothalamus) of a subject atdifferent times, and hybridized to a chip containing oligonucleotideprobes which are complementary to the transcripts of the desired genes,to compare expression of a large number of genes at various stages ofschizophrenia, bipolar disorder and/or ADHD.

In another aspect, an animal model-based assay based on the disclosedgenes can be used to identify agents that can modulate the expression ofone or more genes that are differentially expressed in subjects, e.g.human and/or animal models with schizophrenia, bipolar disorder and/orADHD compared to subjects without schizophrenia, bipolar disorder and/orADHD. Such agents are suitable for use in the treatment ofschizophrenia, bipolar disorder and/or ADHD, and are useful in studiesof the morphogenesis and progression of schizophrenia, bipolar disorderand/or ADHD. These methods generally involve comparing the expressionlevel of one or more of the disclosed genes in a subject, e.g. in asample of the frontal pole and/or hypothalamus that is contacted with acandidate agent to the expression level of the gene or genes in cellsthat are not contacted with the candidate agent. A suitable assay isshown in Example 4 and makes use of the animal model disclosed in thisapplication. Thus, in some embodiments, the method comprises:

-   -   a) contacting the frontal pole and/or hypothalamus of a subject,        e.g. animal model as defined herein with a candidate agent;    -   b) detecting a level of expression of at least one gene        identified in Tables 1, 2 and/or 3 in the frontal pole and/or        hypothalamus (=sample); and    -   c) comparing the level of expression of the gene in the sample        in the presence of the candidate agent with a level of        expression of the gene in the sample that is not contacted with        the candidate agent, wherein a decreased or increased level of        expression in the sample in the presence of the agent relative        to the level of expression in the absence of the agent is        indicative of an agent useful in the treatment of schizophrenia,        bipolar disorder and/or ADHD.

In some embodiments, if expression of only one gene is detected the geneis not any one of the genes of Table 3. The level of expression of thegene is detected by measuring the level of mRNA corresponding to, orprotein encoded by, the gene as described above. As used herein, theterm “candidate agent” refers to any molecule that is capable ofdecreasing the level of mRNA corresponding to, or protein encoded by, atleast one of the disclosed genes. The candidate agent can be natural orsynthetic molecules such as proteins or fragments thereof, smallmolecule inhibitors, nucleic acid molecules, e.g., antisensenucleotides, ribozymes, double-stranded RNAs, organic and inorganiccompounds and the like. Cell-based and/or cell-free assays can also beused to identify compounds which are capable of interacting with aprotein encoded by one of the Genes of the invention or protein-bindingpartner to alter the activity of the protein or its binding partner.Cell-based and/or cell-free assays can also be used to identifycompounds which modulate the interaction between the encoded protein andits binding partner such as a target peptide. In one embodiment,cell-based and/or cell-free assays for identifying such compoundscomprise a cell expressing a protein encoded by any one of the Genes ofthe invention or a reaction mixture containing a protein encoded by oneof the disclosed genes and a test compound or a library of testcompounds in the presence or absence of the binding partner, e.g., abiologically inactive target peptide, or a small molecule. Accordingly,a cell-based and/or cell-free method for identifying agents useful inthe treatment of schizophrenia, bipolar disorder and/or ADHD is providedwhich comprises contacting a protein or functional fragment thereof orthe protein binding partner with a test compound or library of testcompounds and detecting the formation of complexes. For detectionpurposes, the protein can be labeled with a specific marker and the testcompound or library of test compounds labeled with a different marker.Interaction of a test compound with the protein or fragment thereof orthe protein-binding partner can then be detected by measuring the levelof the two labels after incubation and washing steps. The presence ofthe two labels is indicative of an interaction. Interaction betweenmolecules can also be assessed by using real-time BIA (BiomolecularInteraction Analysis, Pharmacia Biosensor AB), which detects surfaceplasmon resonance, an optical phenomenon. Detection depends on changesin the mass concentration of mass macromolecules at the biospecificinterface and does not require labeling of the molecules. In one usefulembodiment, a library of test compounds can be immobilized on a sensorsurface, e.g., a wall of a micro-flow cell. A solution containing theprotein, functional fragment thereof, or the protein-binding partner isthen continuously circulated over the sensor surface. An alteration inthe resonance angle, as indicated on a signal recording, indicates theoccurrence of an interaction. This technique is described in more detailin BIAtechnology Handbook by Pharmacia.

Another embodiment of a cell-free assay comprises:

-   -   a) combining a protein encoded by the at least one gene in        Tables 1, 2 and/or 3, preferentaly at least one gene in Tables 1        and/or 2, the protein binding partner, and a test compound to        form a reaction mixture; and    -   b) detecting interaction of the protein and the protein binding        partner in the presence and absence of the test compounds.

A considerable change (potentiation or inhibition) in the interaction ofthe protein and binding partner in the presence of the test compoundcompared to the interaction in the absence of the test compoundindicates a potential agonist (mimetic or potentiator) or antagonist(inhibitor) of the proteins activity for the test compound. Thecomponents of the assay can be combined simultaneously or the proteincan be contacted with the test compound for a period of time, followedby the addition of the binding partner to the reaction mixture. Theefficacy of the compound can be assessed by using various concentrationsof the compound to generate dose response curves. A control assay canalso be performed by quantitating the formation of the complex betweenthe protein and its binding partner in the absence of the test compound.Formation of a complex between the protein and its binding partner canbe detected by using detectably labeled proteins such as radiolabeled,fluorescently labeled, or enzymatically labeled protein or its bindingpartner, by immunoassay or by chromatographic detection. In preferredembodiments, the protein or its binding partner can be immobilized tofacilitate separation of complexes from uncomplexed forms of the proteinand its binding partner and automation of the assay. Complexation of theprotein to its binding partner can be achieved in any type of vessel,e.g., microtitre plates, micro-centrifuge tubes and test tubes. Inparticularly preferred embodiments, the protein can be fused to anotherprotein, e.g., glutathione-S-transferase to form a fusion protein whichcan be adsorbed onto a matrix, e.g., glutathione sepharose beads (SigmaChemical. St. Louis, Mo.), which are then combined with the labeledprotein partner, e.g., labeled with ³⁵S, and test compound and incubatedunder conditions sufficient to formation of complexes. Subsequently, thebeads are washed to remove unbound label, and the matrix is immobilizedand the radiolabel is determined. The aforementioned cell-free assaysare particularly useful with proteins encoded by the Genes of theinvention. Another method for immobilizing proteins on matrices involvesutilizing biotin and streptavidin. For example, the protein can bebiotinylated using biotin NHS(N-hydroxy-succinimide), using well knowntechniques and immobilized in the well of streptavidin-coated plates.Cell-free assays can also be used to identify agents which are capableof interacting with a protein encoded by the at least one gene andmodulate the activity of the protein encoded by the gene. In oneembodiment, the protein is incubated with a test compound and thecatalytic activity of the protein is determined. In another embodiment,the binding affinity of the protein to a target molecule can bedetermined by methods known in the art.

The present invention also provides for both prophylactic andtherapeutic methods of treating a subject having, or at risk of having,schizophrenia, bipolar disorder and/or ADHD. Subjects at risk for suchdisorders can be identified by a prognostic assay, e.g., as describedabove. Administration of a prophylactic agent can occur prior to themanifestation of symptoms characteristic of schizophrenia, bipolardisorder and/or ADHD, such that development of schizophrenia, bipolardisorder and/or ADHD is prevented or delayed in its progression.Examples of suitable therapeutic agents include, but are not limited to,antisense nucleotides, ribozymes, double-stranded RNAs and antagonists.

Hence, the present invention relates to a method of treating andpreventing schizophrenia, bipolar disorder, and/or ADHD in patient inneed thereof, the method comprising administering to said patient aneffective amount of an agent that can induce a decrease in theexpression of at least one gene with accession number X57514, AI639165,AF064868, AI145494, AA946532, U57500, L17127, L24776, AB020504, U75927,AF096269, AA892801, U49099, AA859597, AA875427, AF028784, L22760,AI014135, AI104513, AA894148, AI073204 and/or D38468; and/or induce anincrease of at least one gene with accession number U20643, H31232,S61973, X85184, J04063, S81353, M33025, AA858621, M74494, AA866358,M64986, AI227715, AA89392, AA859633, AA891969, AI145367, AA893711,Y09000, AA891940, D26564, AI101103, M28648, AF014009, M13100, M59980,M18331, M36418, X57764, U62897, S71570, M16112, AI008131, X12744,AI230260, U48245, H31692, D89863, X06564, M91234, U31554AA875659,AA799421, AI014091, AB012234, U09793, AA800513, AI639381, AA799515,X83546, AI013194, AB008538, X52817, AA893065, H31588, AA859832,AI008074, AA851749, AA894321, AI227608, E13644, AA925762, M72422,M17526, U45479, U86635, AA894089, and/or AA891069; and/or for genesAB016160 and/or U77931 an increase with respect to frontal poleexpression and a decrease to the hypothalamus expression; and/or for thegene U66707 a decrease with respect to the frontal pole and an increasewith respect to the hypothalamus, with the proviso that if expression ofonly one gene is altered that the gene is not any gene of Table 3. Inone embodiment of said method, the agent comprises an isolated nucleicacid molecule comprising an antisense nucleotide sequence derived fromat least one gene with accession number X57514, U66707, AI639165,AF064868, U60578cds, AI145494, AA946532, AB016160, U57500, L17127,M28648, L24776, AB020504, U77931, U75927, AF096269, M892801, U49099,AA859597, AA875427, AF028784, L22760, AI014135, AI104513, AA894148,AI073204 or D38468. In a further embodiment of said method, theantisense nucleotide sequences are derived from at least two genes withaccession number X57514, U66707, AI639165, AF064868, U60578cds,AI145494, AA946532, AB016160, U57500, L17127, M28648, L24776, AB020504,U77931, U75927, AF096269, AA892801, U49099, AA859597, AA875427,AF028784, L22760, AI014135, AI104513, AA894148, AI073204 or D38468. Inanother embodiment of said method, the at least one gene is selectedfrom the group consisting of genes with accessions number U20643,H31232, S61973, AB016160, X85184, J04063, S81353, M33025, X57514,AA858621, M74494, M13100, M59980; AA946532, M18331, M36418, X57764,U62897, U57500, L17127, S71570, M16112, AI008131, M28648, X12744,AI230260, U86635, D38468. In one preferred embodiment of said method,the agent is an antagonist or agonist that inhibits a protein encoded byat least one gene with accession number X57514, U66707, AI639165,AF064868, U60578cds, AI145494, AA946532, AB016160, U57500, L17127,M28648, L24776, AB020504, U77931, U75927, AF096269, AA892801, U49099,AA859597, AA875427, AF028784, L22760, AI014135, AI104513, AA894148,AI073204 or D38468. In another preferred embodiment of said method, theagent is an antagonist or agonist that activates a protein encoded by atleast one gene with accession number U20643, H31232, S61973, AB016160,X85184, J04063, S81353, M33025, AA858621, M74494, AA866358, M64986,AI227715, U77931, AA89392, AA859633, AA891969, AI145367, AA893711,Y09000, AA891940, D26564, AI101103, M28648, AF014009, M13100, M59980,M18331, M36418, X57764, U62897, S71570, M16112, AI008131, X12744,AI230260, U48245, U66707, H31692, D89863, X06564, M91234,U31554AA875659, AA799421, AI014091, AB012234, U09793, AA800513,AI639381, AA799515, X83546, AI013194, AB008538, X52817, AA893065,H31588, AA859832, AI008074, AA851749, AA894321, AI227608, E13644,AA925762, M72422, M17526, U45479, U86635, AA894089 or AA891069.

As used herein, the term “antisense” refers to nucleotide sequences thatare complementary to a portion of an RNA expression product of at leastone of the disclosed genes. “Complementary” nucleotide sequences referto nucleotide sequences that are capable of base-pairing according tothe standard Watson-Crick complementarity rules. That is, purines willbase-pair with pyrimidine to form combinations of guanine:cytosine andadenine:thymine in the case of DNA, or adenine:uracil in the case ofRNA. Other less common bases, e.g., inosine, 5-methylcytosine,6-methyladenine, hypoxanthine and others may be included in thehybridizing sequences and will not interfere with pairing.

When introduced into a host cell, antisense nucleotide sequencesspecifically hybridize with the cellular mRNA and/or genomic DNAcorresponding to the gene(s) so as to inhibit expression of the encodedprotein, e.g., by inhibiting transcription and/or translation within thecell.

The isolated nucleic acid molecule comprising the antisense nucleotidesequence can be delivered, e.g., as an expression vector, which whentranscribed in the cell, produces RNA which is complementary to at leasta unique portion of the encoded mRNA of the gene(s). Alternatively, theisolated nucleic acid molecule comprising the antisense nucleotidesequence is an oligonucleotide probe which is prepared ex vivo and,which, when introduced into the cell, results in inhibiting expressionof the encoded protein by hybridizing with the mRNA and/or genomicsequences of the gene(s).

Preferably, the oligonucleotide contains artificial internucleotidelinkages which render the antisense molecule resistant to exonucleasesand endonucleases, and thus are stable in the cell. Examples of modifiednucleic acid molecules for use as antisense nucleotide sequences arephosphoramidate, phosporothioate and methylphosphonate analogs of DNA asdescribed, e.g., in U.S. Pat. Nos. 5,176,996; 5,264,564; and 5,256,775.General approaches to preparing oligomers useful in antisense therapyare described, e.g., in Van der Krol, BioTechniques, Vol. 6, pp. 958-976(1988); and Stein et al., Cancer Res., Vol. 48, pp. 2659-2668 (1988).

Typical antisense approaches, involve the preparation ofoligonucleotides, either DNA or RNA, that are complementary to theencoded mRNA of the gene. The antisense oligonucleotides will hybridizeto the encoded mRNA of the gene and prevent translation. The capacity ofthe antisense nucleotide sequence to hybridize with the desired genewill depend on the degree of complementarity and the length of theantisense nucleotide sequence. Typically, as the length of thehybridizing nucleic acid increases, the more base mismatches with an RNAit may contain and still form a stable duplex or triplex. One skilled inthe art can determine a tolerable degree of mismatch by use ofconventional procedures to determine the melting point of the hybridizedcomplexes.

Antisense oligonucleotides are preferably designed to be complementaryto the 5′ end of the mRNA, e.g., the 5′ untranslated sequence up to andincluding the regions complementary to the mRNA initiation site, i.e.,AUG. However, oligonucleotide sequences that are complementary to the 3′untranslated sequence of mRNA have also been shown to be effective atinhibiting translation of mRNAs as described, e.g., in Wagner, Nature,Vol. 372, pp. 333 (1994). While antisense oligonucleotides can bedesigned to be complementary to the mRNA coding regions, sucholigonucleotides are less efficient inhibitors of translation.

Regardless of the mRNA region to which they hybridize, antisenseoligonucleotides are generally from about 15 to about 25 nucleotides inlength.

The antisense nucleotide can also comprise at least one modified basemoiety, e.g., 3-methylcytosine, 5-methylcytosine, 7-methylguanine,5-fluorouracil, 5-bromouracil, and may also comprise at least onemodified sugar moiety, e.g., arabinose, hexose, 2-fluorarabinose, andxylulose.

In another embodiment, the antisense nucleotide sequence is analpha-anomeric nucleotide sequence. An alpha-anomeric nucleotidesequence forms specific double stranded hybrids with complementary RNA,in which, contrary to the usual beta-units, the strands run parallel toeach other as described e.g., in Gautier et al., Nucl. Acids. Res., Vol.15, pp. 6625-6641 (1987).

Antisense nucleotides can be delivered to cells which express thedescribed genes in vivo by various techniques, e.g., injection directlyinto the relevant brain tissue site, entrapping the antisense nucleotidein a liposome, by administering modified antisense nucleotides which aretargeted to the relevant brain cells by linking the antisensenucleotides to peptides or antibodies that specifically bind receptorsor antigens expressed on the cell surface.

However, with the above-mentioned delivery methods, it may be difficultto attain intracellular concentrations sufficient to inhibit translationof endogenous mRNA. Accordingly, in a preferred embodiment, the nucleicacid comprising an antisense nucleotide sequence is placed under thetranscriptional control of a promoter, i.e., a DNA sequence which isrequired to initiate transcription of the specific genes, to form anexpression construct. The use of such a construct to transfect cellsresults in the transcription of sufficient amounts of single strandedRNAs to hybridize with the endogenous mRNAs of the described genes,thereby inhibiting translation of the encoded mRNA of the gene. Forexample, a vector can be introduced in vivo such that it is taken up bya cell and directs the transcription of the antisense nucleotidesequence. Such vectors can be constructed by standard recombinanttechnology methods. Typical expression vectors include bacterialplasmids or phage, such as those of the pUC or Bluescript.™ plasmidseries, or viral vectors such as adenovirus, adeno-associated virus,herpes virus, vaccinia virus and retrovirus adapted for use ineukaryotic cells. Expression of the antisense nucleotide sequence can beachieved by any promoter known in the art to act in mammalian cells.Examples of such promoters include, but are not limited to, the promotercontained in the 3′ long terminal repeat of Rous sarcoma virus asdescribed, e.g., in Yamamoto et al., Cell, Vol. 22, pp. 787-797 (1980);the herpes thymidine kinase promoter as described, e.g., In Wagner etal., Proc. Natl. Acad. Sci. USA, Vol. 78, pp. 1441-1445 (1981); the SV40early promoter region as described, e.g., in Bernoist and Chambon,Nature, Vol. 290, pp. 304-310 (1981); and the regulatory sequences ofthe metallothionein gene as described, e.g., in Brinster et al., Nature,Vol. 296, pp. 39-42 (1982).

Ribozymes are RNA molecules that specifically cleave othersingle-stranded RNA in a manner similar to DNA restrictionendonucleases. By modifying the nucleotide sequences encoding the RNAs,ribozymes can be synthesized to recognize specific nucleotide sequencesin a molecule and cleave it as described, e.g., in Cech, J. Amer. Med.Assn., Vol. 260, p. 3030 (1988). Accordingly, only mRNAs with specificsequences are cleaved and inactivated.

Two basic types of ribozymes include the “hammerhead”-type as describedfor example in Rossie et al., Pharmac. Ther., Vol. 50, pp. 245-254(1991); and the hairpin ribozyme as described, e.g., in Hampel et al.,Nucl. Acids Res., Vol. 18, pp. 299-304 (1999) and U.S. Pat. No.5,254,678. Intracellular expression of hammerhead and hairpin ribozymestargeted to mRNA corresponding to at least one of the disclosed genescan be utilized to inhibit protein encoded by the gene.

Ribozymes can either be delivered directly to cells, in the form of RNAoligonucleotides incorporating ribozyme sequences, or introduced intothe cell as an expression vector encoding the desired ribozymal RNA.Ribozyme sequences can be modified in essentially the same manner asdescribed for antisense nucleotides, e.g., the ribozyme sequence cancomprise a modified base moiety.

Double-stranded RNA, i.e., sense-antisense RNA, corresponding to atleast one of the disclosed genes, can also be utilized to interfere withexpression of at least one of the disclosed genes. Interference with thefunction and expression of endogenous genes by double-stranded RNA hasbeen shown in various organisms such as C. elegans as described, e.g.,in Fire et al., Nature, Vol. 391, pp. 806-811 (1998); drosophilia asdescribed, e.g., in Kennerdell et al., Cell, Vol. 95, No. 7, pp.1017-1026 (1998); and mouse embryos as described, e.g., in Wianni etal., Nat. Cell Biol., Vol. 2, No. 2, pp. 70-75 (2000). Suchdouble-stranded RNA can be synthesized by in vitro transcription ofsingle-stranded RNA read from both directions of a template and in vitroannealing of sense and antisense RNA strands. Double-stranded RNA canalso be synthesized from a cDNA vector construct in which the gene ofinterest is cloned in opposing orientations separated by an invertedrepeat. Following cell transfection, the RNA is transcribed and thecomplementary strands reanneal. Double-stranded RNA corresponding to atleast one of the disclosed genes could be introduced into a relevantbrain cell by cell transfection of a construct such as that describedabove.

The term “antagonist” refers to a molecule which, when bound to theprotein encoded by the gene, inhibits its activity. Antagonists caninclude, but are not limited to, peptides, proteins, carbohydrates, andsmall molecules.

In a particularly useful embodiment, the antagonist is an antibodyspecific for the cell-surface protein expressed by the at least one Geneof the invention. Antibodies useful as therapeutics encompass theantibodies as described above. The antibody alone may act as an effectorof therapy or it may recruit other cells to actually effect cellkilling. The antibody may also be conjugated to a reagent such as achemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussistoxin, etc., and serve as a target agent. Alternatively, the effectormay be a lymphocyte carrying a surface molecule that interacts, eitherdirectly or indirectly, with a tumor target. Various effector cellsinclude cytotoxic T cells and NK cells.

Examples of the antibody-therapeutic agent conjugates which can be usedin therapy include, but are not limited to: 1) antibodies coupled toradionuclides, such as ¹²⁵I, ¹³¹I, ¹²³I, ¹¹¹In, ¹⁰⁵Rh, ¹⁵³Sm, ⁶⁷Cu,⁶⁷Ga, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁸⁶Re and ¹⁸⁸Re, and as described, e.g., inGoldenberg et al., Cancer Res., Vol. 41, pp. 4354-4360 (1981);Carrasquillo et al., Cancer Treat. Rep., Vol. 68, pp. 317-328 (1984);Zalcberg et al., J. Natl. Cancer Inst., Vol. 72, pp. 697-704 (1984);Jones et al., Int. J. Cancer, Vol. 35, pp. 715-720 (1985); Lange et al.,Surgery, Vol. 98, pp. 143-150 (1985); Kaltovich et al., J. Nucl. Med.,Vol. 27, p. 897 (1986); Order et al., Int. J. Radiother. Oncol. Biol.Phys., Vol. 8, pp. 259-261 (1982); Courtenay-Luck et al., Lancet, Vol.1, pp. 1441-1443 (1984); and Ettinger et al., Cancer Treat. Rep., Vol.66, pp. 289-297 (1982); (2) antibodies coupled to drugs or biologicalresponse modifiers such as methotrexate, adriamycin, and lymphokinessuch as interferon as described, for, e.g., in Chabner et al., Cancer,Principles and Practice of Oncology, Philadelphia, Pa., J. B. LippincottCo. Vol. 1, pp. 290-328 (1985); Oldham et al., Cancer, Principles andPractice of Oncology, Philadelphia, Pa., J. B. Lippincott Co., Vol. 2,pp. 2223-2245 (1985); Deguchi et al., Cancer Res., Vol. 46, pp.3751-3755 (1986); Deguchi et al., Fed. Proc., Vol. 44, p. 1684 (1985);Embleton et al., Br. J. Cancer, Vol. 49, pp. 559-565 (1984); and Pimm etal., Cancer Immunol. Immunother., Vol. 12, pp. 125-134 (1982); (3)antibodies coupled to toxins, as described, for example, in Uhr et al.,Monoclonal Antibodies and Cancer, Academic Press, Inc., pp. 85-98(1983); Vitetta et al., Biotechnology and Bio. Frontiers, Ed. P. H.Abelson, pp. 73-85 (1984); and Vitetta et al., Science, Vol. 219, pp.644-650 (1983); (4) heterofunctional antibodies, for example, antibodiescoupled or combined with another antibody so that the complex binds bothto the carcinoma and effector cells, e.g., killer cells such as T cells,as described, for example, in Perez et al., J. Exper. Med., Vol. 163,pp. 166-178 (1986); and Lau et al., Proc. Natl. Acad. Sci. USA, Vol. 82,pp. 8648-8652 (1985); and (5) native, i.e., non-conjugated ornon-complexed antibodies, as described in, for example, Herlyn et al.,Proc. Natl. Acad. Sci. USA, Vol. 79, pp. 4761-4765 (1982); Schulz etal., Proc. Natl. Acad. Sci. USA, Vol. 80, pp. 5407-5411 (1983); Caponeet al., Proc. Natl. Acad. Sci. USA, Vol. 80, pp. 7328-7332 (1983); Searset al., Cancer Res., Vol. 45, pp. 5910-5913 (1985); Nepom et al., Proc.Natl. Acad. Sci. USA, Vol. 81, pp. 2864-2867 (1984); Koprowski et al.,Proc. Natl. Acad. Sci. USA, Vol. 81, pp. 216-219 (1984); and Houghton etal., Proc. Natl. Acad. Sci. USA, Vol. 82, pp. 1242-1246 (1985).

Methods for coupling an antibody or fragment thereof to a therapeuticagent as described above are well known in the art and are described,e.g., in the methods provided in the references above. In yet anotherembodiment, the antagonist useful as a therapeutic for treatingschizophrenia, bipolar disorder of ADHD can be an inhibitor of a proteinencoded by one of the disclosed genes. For example, the activity of themembrane-bound serine protease hepsin can be inhibited by utilizingspecific serine protease inhibitors. Such serine-protease inhibitors arewell known in the art as described, e.g., in Leung et al., “ProteaseInhibitors: Current Status and Future Prospects”, J. Med. Chem., Vol.43, pp. 305-341 (2000).

In the case of treatment with an antisense nucleotide, the methodcomprises administering a therapeutically effective amount of anisolated nucleic acid molecule comprising an antisense nucleotidesequence derived from at least one gene identified in Tables 1, 2, or 3with the proviso that if expression of only one gene is inhibited thatthe gene is not any one of Table 3, wherein the antisense nucleotide hasthe ability to decrease the transcription/translation of the at leastone gene. In some embodiments, if expression of only one gene isinhibited the gene is not any one of Table 3. The term “isolated”nucleic acid molecule means that the nucleic acid molecule is removedfrom its original environment (e.g., the natural environment if it isnaturally occurring). For example, a naturally occurring nucleic acidmolecule is not isolated, but the same nucleic acid molecule, separatedfrom some or all of the co-existing materials in the natural system, isisolated, even if subsequently reintroduced into the natural system.Such nucleic acid molecules could be part of a vector or part of acomposition and still be isolated, in that such vector or composition isnot part of its natural environment.

With respect to treatment with a ribozyme or double-stranded RNAmolecule, the method comprises administering a therapeutically effectiveamount of a nucleotide sequence encoding a ribozyme, or adouble-stranded RNA molecule, wherein the nucleotide sequence encodingthe ribozyme/double-stranded RNA molecule has the ability to decreasethe transcription/translation of at least one gene identified in Tables1, 2, or 3, with the proviso that if expression of only one gene isinhibited that the gene is not any gene of Table 3.

In the case of treatment with an antagonist or agonist, the methodcomprises administering to a subject a therapeutically effective amountof an antagonist that inhibits a protein encoded by at least one geneidentified in Tables 1, 2, or 3, with the proviso that if expression ofonly one gene is inhibited that the gene is not any gene of Table 3.

A “therapeutically effective amount” of an isolated nucleic acidmolecule comprising an antisense nucleotide, nucleotide sequenceencoding a ribozyme, double-stranded RNA, or antagonist, refers to asufficient amount of one of these therapeutic agents to treatschizophrenia, bipolar disorder, ADHD. The determination of atherapeutically effective amount is well within the capability of thoseskilled in the art. For any therapeutic, the therapeutically effectivedose can be estimated initially either in cell culture assays, e.g., ofneoplastic cells, or in animal models, usually mice, rabbits, dogs, orpigs. The animal model may also be used to determine the appropriateconcentration range and route of administration. Such information canthen be used to determine useful doses and routes for administration inhumans.

Therapeutic efficacy and toxicity may be determined by standardpharmaceutical procedures in cell cultures or experimental animals,e.g., ED50 (the dose therapeutically effective in 50% of the population)and LD50 (the dose lethal to 50% of the population). The dose ratiobetween toxic and therapeutic effects is the therapeutic index, and itcan be expressed as the ratio, LD50/ED50. Antisense nucleotides,ribozymes, double-stranded RNAs, and antagonists which exhibit largetherapeutic indices are preferred. The data obtained from cell cultureassays and animal studies is used in formulating a range of dosage forhuman use. The dosage contained in such compositions is preferablywithin a range of circulating concentrations that include the ED50 withlittle or no toxicity. The dosage varies within this range, dependingupon the dosage form employed, sensitivity of the patient, and the routeof administration.

The exact dosage will be determined by the practitioner, in light offactors related to the subject that requires treatment. Dosage andadministration are adjusted to provide sufficient levels of the activemoiety or to maintain the desired effect. Factors which may be takeninto account include the severity of the disease state, general healthof the subject, age, weight, and gender of the subject, diet, time andfrequency of administration, drug combination(s), reactionsensitivities, and tolerance/response to therapy.

Normal dosage amounts may vary from 0.1 to 100,000 micrograms, up to atotal dose of about 1 g, depending upon the route of administration.Guidance as to particular dosages and methods of delivery is provided inthe literature and generally available to practitioners in the art.Those skilled in the art will employ different formulations fornucleotides than for antagonists.

For therapeutic applications, the antisense nucleotides, nucleotidesequences encoding ribozymes, double-stranded RNAs (whether entrapped ina liposome or contained in a viral vector) and antibodies are preferablyadministered as pharmaceutical compositions containing the therapeuticagent in combination with one or more pharmaceutically acceptablecarriers. The compositions may be administered alone or in combinationwith at least one other agent, such as stabilizing compound, which maybe administered in any sterile, biocompatible pharmaceutical carrier,including, but not limited to, saline, buffered saline, dextrose, andwater. The compositions may be administered to a patient alone, or incombination with other agents, drugs or hormones.

The pharmaceutical compositions may be administered by any number ofroutes, including, but not limited to, oral, intravenous, intramuscular,intra-articular, intra-arterial, intramedullary, intrathecal,intraventricular, transdermal, subcutaneous, intraperitoneal,intranasal, enteral, topical, sublingual, or rectal means.

In addition to the active ingredients, these pharmaceutical compositionsmay contain suitable pharmaceutically-acceptable carriers comprisingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Furtherdetails on techniques for formulation and administration may be found inthe latest edition of Remington's Pharmaceutical Sciences (MaackPublishing Co., Easton, Pa.).

Pharmaceutical compositions for oral administration can be formulatedusing pharmaceutically acceptable carriers well known in the art indosages suitable for oral administration. Such carriers enable thepharmaceutical compositions to be formulated as tablets, pills, dragees,capsules, liquids, gels, syrups, slurries, suspensions, and the like,for ingestion by the patient.

Pharmaceutical preparations for oral use can be obtained throughcombination of active compounds with solid excipient, optionallygrinding a resulting mixture, and processing the mixture of granules,after adding suitable auxiliaries, if desired, to obtain tablets ordragee cores. Suitable excipients are carbohydrate or protein fillers,such as sugars, including lactose, sucrose, mannitol, or sorbitol;starch from corn, wheat, rice, potato, or other plants; cellulose, suchas methyl cellulose, hydroxypropylmethyl-cellulose, or sodiumcarboxymethylcellulose; gums including arabic and tragacanth; andproteins such as gelatin and collagen. If desired, disintegrating orsolubilizing agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, alginic acid, or a salt thereof, such as sodiumalginate.

Dragee cores may be used in conjunction with suitable coatings, such asconcentrated sugar solutions, which may also contain gum arabic, talc,polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titaniumdioxide, lacquer solutions, and suitable organic solvents or solventmixtures. Dyestuffs or pigments may be added to the tablets or drageecoatings for product identification or to characterize the quantity ofactive compound, i.e., dosage.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a coating, such as glycerol or sorbitol. Push-fit capsulescan contain active ingredients mixed with a filler or binders, such aslactose or starches, lubricants, such as talc or magnesium stearate,and, optionally, stabilizers. In soft capsules, the active compounds maybe dissolved or suspended in suitable liquids, such as fatty oils,liquid, or liquid polyethylene glycol with or without stabilizers.

Pharmaceutical formulations suitable for parenteral administration maybe formulated from aqueous solutions, preferably in physiologicallycompatible buffers such as Hanks' solution, Ringer's solution, orphysiologically buffered saline. Aqueous injection suspensions maycontain substances which increase the viscosity of the suspension, suchas sodium carboxymethyl cellulose, sorbitol, or dextran. Additionally,suspensions of the active compounds may be prepared as appropriate oilyinjection suspensions. Suitable lipophilic solvents or vehicles includefatty oils such as sesame oil, or synthetic fatty acid esters, such asethyl oleate or triglycerides, or liposomes. Non-lipid polycationicamino polymers may also be used for delivery. Optionally, the suspensionmay also contain suitable stabilizers or agents which increase thesolubility of the compounds to allow for the preparation of highlyconcentrated solutions.

For topical or nasal administration, penetrants appropriate to theparticular barrier to be permeated are used in the formulation. Suchpenetrants are generally known in the art.

The pharmaceutical compositions of the present invention may bemanufactured in a manner that is known in the art, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping, or lyophilizing processes.

The pharmaceutical composition may be provided as a salt and can beformed with many acids, including but not limited to, hydrochloric,sulfuric, acetic, lactic, tartaric, malic, succinic, etc. Salts tend tobe more soluble in aqueous or other protonic solvents than are thecorresponding free base forms. In other cases, the preferred preparationmay be a lyophilized powder which may contain any or all of thefollowing: 1-50 mM histidine, 0. 1-2% sucrose, and 2-7% mannitol, at apH range of 4.5 to 5.5, that is combined with buffer prior to use.

After pharmaceutical compositions have been prepared, they can be placedin an appropriate container and labeled for treatment of an indicatedcondition. For administration of the antisense nucleotide or antagonist,such labeling would include amount, frequency, and method ofadministration. Those skilled in the art will employ differentformulations for antisense nucleotides than for antagonists, e.g.,antibodies or inhibitors. Pharmaceutical formulations suitable for oraladministration of proteins are described, e.g., in U.S. Pat. Nos.5,008,114; 5,505,962; 5,641,515; 5,681,811; 5,700,486; 5,766,633;5,792,451; 5,853,748; 5,972,387; 5,976,569; and 6,051,561.

In another aspect, the treatment of a subject with a therapeutic agent,such as those described above, can be monitored by detecting the levelof expression of mRNA or protein encoded by at least one of thedisclosed genes identified in Tables 1 or 2, or the activity of theprotein encoded by the at least one gene. These measurements willindicate whether the treatment is effective or whether it should beadjusted or optimized. Accordingly, one or more of the genes describedherein can be used as a marker for the efficacy of a drug duringclinical trials.

In a particularly useful embodiment, a method for monitoring theefficacy of a treatment of a subject having schizophrenia, bipolardisorder and/or ADHD, or at risk of. or having schizophrenia, bipolardisorder and/or ADHD with an agent (e.g., an antagonist, protein,nucleic acid, small molecule, or other therapeutic agent or candidateagent identified by the screening assays described herein) is providedcomprising:

-   -   a) obtaining a pre-administration sample from a subject prior to        administration of the agent,    -   b) detecting the level of expression of mRNA corresponding to,        or protein encoded by the at least one gene, or activity of the        protein encoded by the at least one gene identified in Tables 1        or 2 in the pre-administration sample;    -   c) obtaining one or more post-administration samples from the        subject,    -   d) detecting the level of expression of mRNA corresponding to,        or protein encoded by the at least one gene, or activity of the        protein encoded by the at least one gene in the        post-administration sample or samples,    -   e) comparing the level of expression of mRNA or protein encoded        by the at least one gene, or activity of the protein encoded by        the at least one gene in the pre-administration sample with the        level of expression of mRNA or protein encoded by the at least        one gene, or activity of the protein encoded by the at least one        gene in the post-administration sample or samples, and    -   f) adjusting the administration of the agent accordingly.

In some embodiments, if expression corresponding to, or activity ofprotein encoded by, only one gene is detected the gene is not any geneof Table 3. In other embodiments, if expression corresponding to, oractivity of protein encoded by, only one gene is detected the gene isnot any gene of Table 3. For example, increased administration of theagent may be desirable to decrease the level of expression or activityof the at least one gene to lower levels than detected, i.e., toincrease the effectiveness of the agent. Alternatively, decreasedadministration of the agent may be desirable to increase expression oractivity of the at least one gene to higher levels than detected, i.e.,to decrease the effectiveness of the agent.

In another aspect, a viral vector is provided which comprises a promoterand/or an enhancer or other regulatory element of a gene selected fromthe group consisting of at least one of the genes identified in Tables1, 2, or 3 operably linked to the coding region of a gene that isessential for replication of the vector, wherein the vector is adaptedto replicate upon transfection into a diseased cell. The promotersequences can be discerned by searching the publicly available databasesfor BAC clones that cover the entire gene; thereafter, the cDNA for thegene can be compared to the genomic sequence. This will generally revealthe intron-exon boundaries and the start site of the gene. Once theseare established, the promoter sequences can be inferred. Such vectorsare able to selectively replicate in a brain cell, e.g. frontal lobeand/or hypothalamus, of a patient having schizophrenia, bipolar disorderand/or ADHD, but not in a non-diseased cell. The replication isconditional upon the presence in a diseased cell, and not in anon-diseased (=standard expression profile) cell, of positivetranscription factors that activate the promoter of the disclosed genes.It can also occur by the absence of transcription inhibiting factorsthat normally occur in a non-diseased cell and prevent transcription asa result of the promoter. Accordingly, when transcription occurs, itproceeds into the gene essential for replication, such that in thediseased cell, but not in non-diseased cell, replication of the vectorand its attendant functions occur. With this vector, a diseased cell,e.g., a brain cell, e.g. hypothalamus and/or frontal lobe of a patienthaving schizophrenia, bipolar disorder and/or ADHD, can be selectivelytreated, with minimal systemic toxicity.

In one embodiment, the viral vector is an adenoviral vector, whichincludes a coding region of a gene essential for replication of thevector, wherein the coding region is selected from the group consistingof E1a, E1b, E2 and E4 coding regions. The term “gene essential forreplication” refers to a nucleic acid sequence whose transcription isrequired for the vector to replicate in the target cell. Preferably, thegene essential for replication is selected from the group consisting ofthe E1A and E1b coding sequences. Particularly preferred is theadenoviral E1A gene as the gene essential for replication. Methods formaking such vectors are well know to the person of ordinary skill in theart as described, e.g., in Sambrook et al., in Molecular Cloning: ALaboratory Manual, Cold Spring Harbor, N.Y., 1989.

In a further embodiment, the invention provides nucleic acid constructsin which a heterologous gene product is expressed under the control of apromoter and/or an enhancer or other regulatory element of a geneselected from the group consisting of at least one of the genesidentified in Tables 1, 2, 3, 5, 6 or 7. Such heterologous gene productsare expressed when the construct is present in diseased cells, but notin normal, non-diseased cells. The heterologous gene product provides,in some embodiments, for the inhibition, prevention, or destruction ofthe growth of the diseased cell. The gene product can be RNA, e.g.,antisense RNA or ribozyme, or proteins such as a cytokine, e.g.,interleukin, interferon, or toxins such as diphtheria toxin, pseudomonastoxin, etc. The heterologous gene product can also be a negativeselective marker such as cytosine deaminase. U.S. Pat. No. 6,057,299,for example, describes the construction and use of nucleic acidconstructs in which heterologous genes are placed under the control of aPSA enhancer. The nucleic acid constructs can be introduced into targetcells by methods known to those of skill in the art. For example, onecan incorporate the constructs into an appropriate vector such as thosedescribed above.

The vector of the present invention can be transfected into a helpercell line for viral replication and to generate infectious viralparticles. Alternatively, transfection of the vector or other nucleicacid into a nerve cell can take place by electroporation, calciumphosphate precipitation, microinjection, or through liposomes, includingproteoliposomes.

In a further embodiment, the invention provides a novel heuristic animalmodel for schizophrenia, bipolar disorder and/or ADHD as shown inexample 1.

The following examples are included to demonstrate preferred embodimentsof the invention.

EXAMPLE 1 Animal Model

A) The repeated variable prenatal stress paradigm: Timed pregnantSprague Dawley rats (Charles River Laboratories, USA) are subjected to avariable repeated stress during the last week of the gestation, startingon embryonic day (E) 14 and continuing until the natural delivery of thepups at E22. The stress paradigm consists of a 1 h restraint in thecylindrical restrainers (×3/day), exposure to a cold environment (+4°C., 6 h), over night food deprivation, swim stress (×3/day) and reversalof the light-dark cycle. All of the mothers are exposed to the samestressors but in a randomized manner to prevent accomodation and, thesame stresses are applied at the same time of the day. During theprenatal stress period, all stresses are performed at least twice.Control dams remain in the animal room and are not exposed to anyunusual procedures. Following the delivery, the dam and her pups areleft undisturbed in their cages until weaning on the postnatal day (P)21, when the male and the female offspring are separated and housed with1-2 littermates per cage with free access to food and water.

B) The acute stress procedure: On the morning of P56 (at 0900 h), afteran acclimation period of 5-7 d in the laboratory environment tore-establish homeostasis and basal levels of gene expression after aconsiderable release of corticosterone induced by moving the animalsfrom animal room to the laboratory (The laboratory environment issoundproof, and has its own ventilation system. An acclimation period ofmin. 48 h was required to re-establish homeostasis and basal levels ofgene expression J. Koenig, unpublished observation.), the prenatallystressed offspring and the prenatally non-stresssed controls are placedin cylindrical plastic restrainers for 30 min. At the end of the acuterestraint session, the animals were either sacrificed by decapitation orreturned to their cages to be sacrificed at 120 min or 24 h afterremoval from the restraint device. Rats designated to the baseline groupto establish the basal corticosterone levels and gene expression aresacrificed without exposure to acute stress throughout the course of theexperiment to control for circadian differences between the animals.

C) Behavioral assessment: Behavioral assessments consisting of basal andpsychomotor stimulant-induced locomotor activity, prepulse inhibition(PPI; a measure of sensorimotor gating; Loss of normal PPI has beenthought to be a measure of sensorymotor gating and has been shown to bedisrupted in schizophrenia; Braff and Geyer, Arch Gen Psychiatry.47:181-188 (1987); Geyer and Braff, Schizophr Bull. 13:643-668 (1990)),N40 (auditory evoked potential, measure of sensory gating in rats;Bickford-Wimer et al., Biol Psychiatry. 27:183-192 (1990)) and acutestress reactivity are done on the male offspring on postnatal days 35and 56. Control animals are obtained from dams not exposed to stressduring gestation. Prenatal stress exposure induce a heightened locomotorresponse to both amphetamine and phencyclidine only on post-natal day56. Moreover, sensorimotor gating assayed by both PPI and N40 aredisrupted in animals exposed to prenatal stress. Deficits inglucocorticoid feedback are noted in animals exposed to prenatal stresson post-natal day 56. Plasma corticosterone levels are measured inprenatally stressed and the non-stressed controls at baseline and, 30min., 2 h and 24 h after acute stress exposure. The prenatally stressedshow a prolonged elevation of corticosterone after the acute stress.

The animals used in this study are maintained in fascilities fullyaccredited by the American Association for the Accreditation oflaboratory Animal care (AAALAC) and the studies are conducted inaccordance with the Guide for Care and Use of Laboratory Animalsprovided by the NIH.

EXAMPLE 2 Identification of Potential Diagnostic Markers and TherapeuticTargets

A) Probe preparation and microarray hybridization: The brains areremoved and frontal pole and hypothalamus are quickly dissected, placedin finely powdered dry ice till the samples are frozen and then wrappedin aluminium foil. For dissection of the frontal pole, the brain isplaced on the dorsal side and a clean razor blade is used to make afrontal cut beginning at the rostral pole of the olfactory tubercle andproceeding through the dorsal cortex. For hypothalamus, the brain isplaced on the dorsal side and a clean razor blade is used to make avertical cut at the rostral tip of the optic chiasm and behind themammillary bodies. Cuts on the lateral aspects are made in thehippocampal sulci. The cuts are defined by the Circle of Willis. Oncethis piece of tissue is isolated, it is cut at a thickness of 1.5 mmwhich contains all the hypothalamic nuclei and thus is withoutcontamination by the thalamus or other brain region. The samples arestored at −80° C. until isolation of the total RNA.

Total RNA is isolated using peqGOLD RNAPure™ (PEQLAB BiotechnologieGmbH, Germany), total RNA is DNAse treated (10 U RNAse free DNAse 2U/μl; Ambion and 1 U/μl of the final volume RNAse inhibitor SuperasIn;20 U/μl; Ambion; USA) and repurified using RNeasy (Qiagen Inc., USA).Thereafter, the samples are labeled and hybridized individually on ratgenome RG-U34A microarrays (Affymetrix, Santa Clara, Calif., USA) aspreviously described (Lockhart et al., Nat Biotechnol. 14:1675-1680(1996); Affymetrix Genechip® Expression Analysis Technical Protocol(2001) Affymetrix, Inc., USA). Primary image analysis of the arrays isperformed using the Genechip 3.2 software package (Affymetrix, SantaClara, Calif., USA) and images are scaled to an average hybridizationintensity (average difference value) of 150. For the frontal polesamples (n=48) the background, the percentage of present values, theAFFX-GAPDH3′/AFFX-GAPDH5′ probe expression ratio and the AFFX-GAPDH3′probe variation are 62.5±9.45, 40.89±3.01, 6.86±7.36 and 23%respectively and, for the hypothalamus samples (n=35) 69.61±9.81,44.78±2.54, 2.07±0.99 and 11%, which indicates that the gene chips areof good quality and the variation is relatively small, as compared toother studies performed using similar method and material.

B) Validation of the animal model: Hierarchical clustering of theexperiment based on Pearson correlation around zero of the geneexpression of all 8799 genes on the Affymetrix RGU34A microarray isperformed for the frontal pole samples. Each timepoint (baseline (BL),24 h, 30 minutes, 2 h) reflects the average expression of six individualsamples in that group. The non-stressed controls and the prenatallystressed clustered separately. In both main treatment groups, the globalgene expression at the 2 h timepoint after acute stress differs mostfrom the BL situation. The gene expression returns close to the baseline24 h after the acute stress in the prenatally non-stressed animals.However, in the prenatally stressed, the gene expression at 24 h afteracute stress resembled more closely the 30 min or the 2 h situation thanthe BL situation thus reflecting a prolonged stress response at theglobal gene expression level in the prenatally stressed animals. Thismolecular pattern thus goes well and fits well with the behavior changesobserved in the animals of example 1.

B) Microarray analysis: Microarray analysis is performed usingGeneSpring® 4.2.1 (Silicon Genetics, Redwood City, Calif., USA) andNovartis Pharmacogenetics Gene Expression Analysis Tools (fordescription see U.S. Ser. No. 60/377,446). Three different normalizationapproached are followed by using raw data, the minimum change of geneexpression and a statistical restriction exercise (see below) to arriveat a list of consistently differentially expressed genes that occurredin common in at least two separate comparisons based on differentnormalizations. The normalization approaches used are:

Approach 1) The expression value for each gene on a chip is divided bythe mean of all expression values on that chip assuming that this is atleast 10. The bottom 10% is used as a test for correct backgroundsubtraction. Thereafter, each gene is normalized to itself by dividingall measurements for that gene by the median of the gene's expressionvalues over all the samples. Only genes having a raw data expressionvalue ≧100 at least in one of the two conditions (non-stressed vs.stressed) are further analysed. Furthermore, genes showing a minimumfold difference of 1.5 in the expression between the non-stressed andthe stressed animals with a statistical significance p≦0.005 (Wilcoxontest) are considered as differentially expressed genes, i.e. 29 genes inthe frontal lobe and 82 genes in the hypothalamus fulfill this criteria.

Approach 2) The expression value for each gene on a chip is divided bythe mean of all expression values on that chip. Thereafter, GAPDH 3′expression on each chip is divided by the average of GAPDH 3′ expressionacross the whole sample set. Result of the GAPDH 3′/Avg GAPDH 3′ is thenused to divide the expression values for other genes on that chip. Theprocedure brings the GAPDH 3′ expression value on every chip to the samevalue, and thus normalizes the global gene expression with respect toGAPDH 3′. Again, only genes having raw data expression values ≧100 atleast in one of the two conditions (non-stressed vs. stressed) arefurther selected. Thereafter, genes showing a minimum fold difference of1.5 in the expression between the non-stressed and the stressed animalswith a statistical significance p≦0.005 (Wilcoxon test) are consideredas differentially expressed, i.e. 42 genes in the frontal lobe and 139genes in the hypothalamus fulfill this criteria.

Approach 3) The signal intensity on each chip has been scaled to 150(arbitrary intensity based on experience), thus no normalization isapplied to this set of data. Genes are selected based on logarithmicaverage expression difference between the study groups. Genes showing aminimum fold difference of 1.5 in the expression between thenon-stressed and the stressed with a statistical significance p≦0.005(Wilcoxon test) are considered as differentially expressed, i.e. 75genes in the frontal lobe and 100 genes in the hypothalamus fulfill thiscriteria.

Thus, only genes that are identified as being differentially expressedin more than one approach shown above are included on a list ofconstantly differentially expressed genes, i.e. 31 genes in the frontalpole (see Table 1) and 66 genes (see Table 2) in the hypothalamusfulfill this final criteria.

Arbitrary expression levels of Genes of the invention are shown todemonstrate the differential levels of expression of said genes as shownin Table 1 and 2 below: TABLE 1 Transcript Levels in the frontal pole ofprenatally stressed rats compared to non stressed rats Accession no.Gene name Up/down regulated U20643 Aldolase A down H31232 Cox IV-2 downS61973 NMDA receptor glutamate-binding subunit (grina) down AB016160GABA-B receptor 1c down X85184 Ras-related GTPase, ragB down J04063CamKII gamma down S81353 Sulfated glycoprotein-1; Prosaposin down M33025Parathymosin down X57514 GABA(A) receptor gamma-1 subunit up AA858621CamKII inhibitor alpha down AA866358 EST, similar to 2310042M24; bysynteny CGI-31 down U66707 Densin-180 up M64986 Rat amphoterin/Hmgb1(high motility group box 1) down AI227715 Retinoblastoma-like 2 downU77931 Ribin down AI639165 cDNA; 94% similar to FLJ11753 up AA89392 4Kruppel-like factor 13 down AA859633 EST, similar to PD2 protein downAA891969 Nuclear DNA-binding protein (C1D) down AI145367 Cap2, adenylylcylase-associated protein 2 down AF064868 Begain up AA893711 EST similarto HS44 mouse; Dnaj (Hsp40) down Y09000 Dendrin down AA891940 EST highlysimilar to RHOC (ras homolog 9) down D26564 RATCDS37, similar to cdc37down AI101103 Vamp2 (synaptobrevin II) down M74494 Na, K-ATPase alpha-1subunit down AF014009 Acidic calcium-independent phospholipase A2 downAI145494 Synapsin 2 up(Notes:Accession number can be used to identify the unique identity of eachgene at NCBI - UniGene at http://www.ncbi.nlm.nih.gov/UniGene/;“up/down regulation” indicates that the genes is differentiallyexpressed in the frontal pole# according to the criteria laid down in example 2, i.e. an at least 1.5fold difference in the expression between the non-stressed and thestressed animals with a statistical significance of p ≦ 0.005 (Wilcoxontest);i.e. “up” indicates an up regulation whereas “down” indicates a downregulation)

TABLE 2 Transcript Levels in the hypothalamus of prenatally stressedrats compared to non-stressed rats Up/down Accession no. Gene nameregulated M13100 Heme oxygenase-3 (HO-3) down M59980 Voltage-gated K+channel protein (RK5) down AA946532 Abcd3 up M18331 Protein kinase Cepsilon down M36418 Glutamate receptor (GluR-A, gria1, glur1) downAB016160 GABAB receptor 1c up X57764 ET-B endothelin receptor downU62897 Carboxypeptidase D precursor (Cpd) down U57500 Protein tyrosinephosphatase alpha up L17127 psmb4; proteasome (prosome, macropain) upS71570 CaMKII gamma-b down M16112 CaMKII beta down AI008131S-adenosylmethionine decarboxylase down M28648 Na, K-ATPase alpha-2 upX12744 c-erb-A thyroid hormone receptor down AI230260 casein kinase IIbeta down U48245 PKC-binding protein Nel down U66707 Densin-180 downH31692 eif2C2 (Gerp95) down D89863 M-Ras down X06564 NCAM down M91234VL30 element down U31554 Limbic system-associated membrane down proteinL24776 Tropomyosin non-muscle isoform NM3 up AB020504 PMF31 up U77931Ribin up U75927 Cytochrome oxidase subunit VIIa up AF096269 epsin 2 upAA892801 Eef2 up U49099 Golgi SNAP receptor complex 1 (Gosr1) upAA875659 alpha internexin down AA859597 ESTs, similar to Kcnk6 upAA875427 Aes up AA799421 ESTs, similar to PKC epsilon down AI014091Cited 2 down AB012234 NF1-X1 down U09793 p21 (c-Ki-ras); kras2 downAA800513 ESTs; similar to AI481500 and K08H10 down AF028784 GFAP alphaand delta up AI639381 ESTs, similar to Ras suppressor 1 down AA799515ESTs, similar to WSB-2 down X83546 Leucocyte common antigen-related(lar2) down AI013194 elF-5 down AB008538 alcam down X52817 C1-13 geneproduct; reticulon 1 down AA893065 ESTs, similar to NPEPPS down H31588ESTs, similar to 2210408F11 gene down AA859832 ESTs, similar to AK004010and down AP001458.5 AI008074 ESTs, similar to HSP90 beta down AA851749Sfrs10 down AA894321 ESTs, similar to FENS1 down AI227608microtubule-associated protein tau down L22760 GATA-GT1 (gata6) upAI014135 beta-carotene 15 up AI104513 cytochrome c oxidase subunit Va upAA894148 ESTs, similar to ApoA-IV up AI073204 Ywhae up E13644 Neurodap-1down AA925762 Marcs down M72422 Glutamic acid decarboxylase (GAD65) downM17526 GTP-binding protein (G-alpha-0) down U45479 Synaptojanin; synj1down U86635 Glutathione S-transferase M down D38468 BIT up AA894089Neurodegeneration-associated protein 1 down AA891069 ESTs, similar toSFRS protein kinase 2 down(Notes:Accession number can be used to identify the unique identity of eachgene at NCBI - UniGene at http://www.ncbi.nlm.nih.gov/UniGene/;“up/down regulation” indicates that the genes is differentiallyexpressed in the hypothalamus according to the criteria laid down inexample 2, i.e. an at least 1.5 fold difference in the expressionbetween the non-stressed# and the stressed animals with a statistical significance of p ≦ 0.005(Wilcoxon test);i.e. “up” indicates an up regulation whereas “down” indicates a downregulation)

TABLE 3 Genes identified in Table 1 and 2 which are known to beassociated with Schizophrenia, ADHD and/or bipolar disorder Accessionno. Gene name Reference AI101103 Vamp2 (synaptobrevin II) Sokolov etal., (2000) Biol. Psych. 48: 184-196. 196. M72422 Glutamic aciddecarboxylase (GAD65) Hakak et al., (2001) Proc. Nat. Acad. Sci., 98:4746-4751. M17526 GTP-binding protein (G-alpha-0) Tani et al., (2001)Mol. Psychiatry 6: 359. AF014009 acidic calcium-independentphospholipase Ross et al. (1997) A2 (aiPLA2) Arch. Gen. Psych. 54:487-494. Ross et al (1999) Brain Res. 821: 407-413. U45479 Synaptojanin;synj1 Saito et al. (2001) Mol Psychiatry 6: 387-395. X06564 NCAM Barbeauet al.(1995) Proc. Natl. Acad. Sci. USA 92: 2785-2789 AI145494 Synapsin2 Mirnics et al., (2000) Neuron 28: 53-67.

EXAMPLE 3 Real-Time (RT) PCR: Confirmation of Differentially RegulatedGenes

Nine genes from the 35 most constantly differentially expressed in thefrontal pole are chosen based on their drugability and validation value(e.g. genes that are already known to be associated e.g. withschizophrenia (see Table 3)) for quantitation using a fluorescence-basedreal time PCR (Taqman, Applied Biosystems, Foster City, Calif.; Gibsonet al., 1996; Heid et al., 1996). The following primers and probes(Microsynth, Balgach, Switzerland) are designed using ABI PrimerExpresssoftware: TABLE 4 The sequence of the probe pairs for RT-PCR: SEQ PrimerName Sequence 5′-3′ ID NO: AiPLA2- CAGCTGCAGTTCCGTAGAAAGA 1 Forward(F)AiPLA2- GCGAGCGACCTACGCG 2 Reverse(R) AiPLA2-ProbeTGTGGCGTGGTCACAGCCGAAG 3 (TQ) Aldolase A-F AGGAAGAGGCATCCATCAACC 4Aldolase A-R GAAAGTCAAGGCCCATGGC 5 Aldolase A-TQCAATGCTATCAACAAGTGTCCCCTGCTGA 6 Densin 180-F GCCTTGACCACCCTGGAAA 7Densin 180-R GCTCCGCTGGAACTGATACAG 8 Densin 180-TQ TAATCACGGCGTTTGCGCGGT9 GabaB1c-F TGGAGAGCTTTGGTCTTTTGC 10 GabaB1c-R GGGTGAGTCACCGCCTACAG 11GabaB1c-TQ TGAGTAGTGATGTTCAGCGGAGGGCC 12 Grina-F CAGCTCAGGTGGCATGGTG 13Grina-R GGAAGCACAGATGGCAATGTC 14 Grina-TQ CTCAGACCCATGCCCCTGCCA 15PGK1-F GCCAACTCGGTTGTGCTTATG 16 PGK1-R GTACTTGTCGGGCATGGGC 17 PGK1-TQCCACCTGGGCCGTCCTGATGGT 18 Ribin-F CAGAGGCTGTTCACCTTGGAG 19 Ribin-RAGGGTGTAAATCTCGCGCC 20 Ribin-TQ TGCTGCGGATATGGGTACGGCC 21 Synapsin2-FACAGATGTCCAAGCCCCCA 22 Synapsin2-R CGCCATGTCAGACCGTACAA 23 Synapsin2-TQACATCTCAGAGCAAGCGTCGACCCAG 24 Vamp2-F CTGCCTGATCTGCTGCCTC 25 Vamp2-RCACCCCTCCTCAAAGAACCA 26 Vamp2-TQ ACCAGGAGAACTGGAGGCTGACCACA 27

All the Taqman probes are 5′FAM (6-carboxyfluorescein)- and 3′ TAMRA(6-carboxytetramethylrhodamine)-labeled. BLAST searches are performed toconfirm primer and probe specificity. Baseline (n=12) and 2 h timepoint(n=11) samples are chosen as template. The amount of total RNA isdetermined using RiboGreen (Molecular Probes, Eugene, Oreg., USA) RNAquatitation method. 400 ng of the total RNA (15 ng/μl) is treated withDNAse I mix (RNAse free DNAse kit, Qiagen, USA). 5 ng of theDNAse-treated RNA is used to control for genomic DNA-contamination usingthe real time PCR and if negative, 250 ng of the RNA is reversetranscribed using random priming and Omniscript RT Kit (Qiagen, USA) asdescribed by the manufacturer. 5 ng of the resulting cDNA is used as atemplate for the following PCR reaction for which the conditions areoptimized so that primers could be used at 300 nM and the probes at 175nM concentration with qPCR Mastermix (Eurogentec, Seraing, Belgium) in a25 μl reaction. The thermal cycle conditions used are as follows: 2 min50° C., 10 min 95° C., followed by 40 two-step cycles at 95° C. for 15 sand one min at 60° C. The relative standard curve method (User Bulletin2, PE Applied Biosystems, 1997) is used to determine the amount of mRNAof the gene of interest relative to an endogenous control gene.Phosphoglycerate kinase I (pgk1) is chosen as an endogenous control genebecause it shows the smallest intersample variation and response toacute stress in our microarray data set (data not shown). For each gene,the samples are run as triplicates or duplicates, and the reactions arerepeated at least once. The reactions for the gene of interest and forthe endogenous control are always performed on the same reaction plate.

EXAMPLE 4 Validation of the Differentially Expressed Genes in the AnimalModel by Known Drugs to Schizophrenia, ADHD and Bipolar Disorder

Timely impregnated Sprague Dawley rats (Charles River Laboratories, USA)are subjected to a variable repeated stress during the last week of thegestation, starting on embryonic day (E) 14 and continuing until thenatural delivery of the pups at E22. The stress paradigm consists of a 1h restraint in the cylindrical restrainers (×3/day), exposure to a coldenvironment (+4° C., 6 h), over night food deprivation, swim stress(×3/day) and reversal of the light-dark cycle. All of the mothers areexposed to the same stressors but in a randomized manner to preventaccomodation and, the same stresses are applied at the same time of theday. During the prenatal stress period, all stresses are performed atleast twice. Control dams remain in the animal room and are not exposedto any unusual procedures. Following the delivery, the dam and her pupsare left undisturbed in their cages until weaning on the postnatal day(P) 21, when the male and the female offspring are separated and housedwith 1-2 littermates per cage with free access to food and water.

On postnatal day (P56) the prenatally stressed and the non-stressedcontrols are divided into corresponding treatment groups and theirbaseline sensory motor gating of is examined using prepulse inhibition(PPI) paradigm. The treatment groups are as follows:

Acute Treatment:

1) Prenatally stressed: Haloperidol i.p. acute dosing regime.

2) Non-stressed controls: Haloperidol i.p., acute dosing regime.

3) Prenatally stressed: Clozapine i.p. acute dosing regime.

4) Non-stressed controls: Clozapine i.p. acute dosing regime.

5) Prenatally stressed: vehicle i.p. (acute dosing control).

6) Non-stressed controls: vehicle i.p. (acute dosing control).

7) Prenatally stressed, methylphenidate i.p. acute dosing regime.

8) Non-stressed controls, methylphenidate i.p. acute dosing regime.

Immediately after acute dosing, the animals are tested for their PPI andsacrificed thereafter. Their brains are dissected, frontal pole,hypothalamus, hippocampus and brain stem dissected and processed forgene expression analysis using microarrays (Affymetrix, USA) and realtime quantitative PCR (Taqman; Applied Biosystems, USA).

Chronic Treatment:

On postnatal day (P56) the prenatally stressed and the non-stressedcontrols are divided into corresponding treatment groups. Their baselinesensory motor gating of is examined using prepulse inhibition (PPI)paradigm. The treatment groups are as follows:

1) Prenatally stressed: Haloperidol p.o. chronic dosing regime.

2) Non-stressed controls: Haloperidol p.o., chronic dosing regime.

3) Prenatally stressed: Clozapine p.o. chronic dosing regime.

4) Non-stressed controls: Clozapine p.o. chronic dosing regime.

5) Prenatally stressed: Fluoxetine p.o. chronic dosing regime.

6) Non-stressed controls: Fluoxetine p.o. chronic dosing regime.

After the treatment, the sensory motor gating is being tested, theanimals will be sacrificed, their brains dissected and selected brainregions will be processed for gene expression analysis using microarrays(Affymetrix, USA) or Taqman (Applied Biosystems, USA).

Prenatally stressed animals have been demonstrated previously to havelearning and memory deficits (Lemaire et al., Proc. Natl. Acad. Sci. USA97:11032-11037 (2000)). To further define the cognitive component of theinitial gene chip finding, a separate group of animals will be subjectedto memory and learning testing (Morris water-maze) and thereaftertreated acutely with rivastigmine, methylphenidate and amphetamine. Thetreatment groups are as follows:

1) Prenatally stressed, rivastigmine i.p.

2) Non-stressed controls, rivastigmine i.p.

3) Prenatally stressed, methylphenidate i.p.

4) Non-stressed controls, methylphenidate i.p.

5) Prenatally stressed, amphetamine i.p.

6) Non-stressed controls, amphetamine i.p.

7) Prenatally stressed, vehicle control i.p.

8) Non-stressed controls, vehicle control i.p.

After the treatment, the cognitive performance is re-assessed.Thereafter, the animals are sacrificed, their brains are dissected andprocessed for microarray (Affymetrix, USA) and Taqman (AppliedBiosystems, USA) analysis of gene expression.

EXAMPLE 5 Identification of the Differential Acute Stress Response inthe Prenatally Stressed Vs. Prenatally Non-Stressed Frontal Pole Samples

Microarray Analysis

Microarray analysis is performed using GeneSpring® 4.2.1 (SiliconGenetics, Redwood City, Calif., USA) and Novartis Pharmacogenetics GeneExpression Analysis Tools (Novartis proprietary). Microarray quality isfirst examined and outliers identified based on the percentage ofpresent values, the background noise level and clustering of theindividual 48 microarrays using hierarchical clustering of all 8799genes on the Affymetrix RG U34 A microarray based on Pearson correlationaround zero (standard correlation with minimum distance of 0.001 andseparation ratio of 0.5). After removal of the outlier microarrays, 25NS controls and 23 PNS microarrays enter the analysis. For the frontalpole samples (n=5-7 per per each group sacrificed at baseline and 30min, 150 min and 24 h after the beginning of the acute stress session;total n=48) the background, the percentage of present values, theAFFX-GAPDH3′/AFFX-GAPDH5′ probe expression ratio and the AFFX-GAPDH3′probe variation are 62.5±9.45, 40.89±3.01, 6.86±7.36 and 23%,respectively. Well above the average variation, a 1.5 fold (=50%) changein the expression signal intensity is chosen to represent a minimum geneexpression change resulting from the acute stress exposure (with rawdata minimum of 100 and p<0.05 (Welch)).

Data Normalization and Analysis

The expression value for each gene on a chip is divided by the mean ofall expression values on that chip assuming that this is at least 10.The bottom tenth percentile is used as a test for correct backgroundsubtraction. Thereafter, each gene is normalized to itself by dividingall measurements for that gene by the median of the gene's expressionvalues over all the samples.

Following the normalization, only genes having raw data expressionvalues ≧100 at least in one of the two conditions (NS and PNS) areincluded in the analysis. Genes showing a minimum fold difference of 1.5in the average expression signal intensity between the NS and the PNSwith a statistical significance of p≦0.005 (Welsch test) are consideredas differentially expressed.

The effect of acute stress on the normal, prenatally non-stressedcontrol male offspring (NS), and the prenatally stressed male offspring(PNS) is studied using two analysis approaches (FIG. 1). In the firstone, the gene expression changes (raw data expression >100, ≧1.5 foldchange, p<0.05) from the baseline to 30 min, 150 min and 24 h followingthe 30 min acute restraint stress are individually analyzed for the NSand the PNS groups. The differentially expressed gene groups consistingof the acute stress response for the NS or PNS individually are thencompared in order to identify a group of common stress response genesand groups of genes that showed a differential expression patternfollowing the acute stress in either the NS or the PNS group.

In the second approach, gene expression at the baseline and individualtimepoints after the acute stress are compared between the NS and PNSanimals and differentially expressed genes (raw data expression >100,≧1.5 fold change, p<0.05) a identified. TABLE 5 Genes constituting the“normal stress response” after exposure to acute stress. AccessionDescription Common genes upregulated from baseline to 30 min after acutestress in the normal controls (prenatally non-stressed) and inprenatally stressed AF030086 Activity and neurotransmitter-induced earlygene 1 (ania-1) AF050659 Activity and neurotransmitter-induced earlygene 7 (ania-7) L01624 Serum and glucocorticoid-regulated kinase (sgk)L26292 FSH-regulated protein mRNA M60921 PC3 NGF-inducibleanti-proliferative putative secreted protein (PC3) M65149CCAAT/enhancerbinding, protein (C/EBP) delta AA891041 jun Bproto-oncogene AA944156 B-cell translocation gene 2, anti-proliferative(Btg2) AI175959 Avian sarcoma virus 17 (v-jun) oncogene homolog (Jun)S81478 CL100 PTPase = oxidative stress-inducible protein tyrosinephosphatase U02553 Dual specificity protein phosphatase 1 U17254Immediate early gene transcription factor NGFI-B, Nr4a1 U19866 Growthfactor Arc U78102 krox20 (egr-2) X06769 c-fos X63594 RL/IF-1 (IkappaB)Common genes upregulated from baseline to 2 h after acute stress D28110MOBP (myelin-associated oligodendrocytic basic protein) Common genesupregulated by acute stress from baseline to 24 h after acute stressJ03179 D-binding protein AF035955 Kinesin-related protein KRP6 (KRP6)Common genes downregulated by acute stress baseline to 30 min afteracute stress X67250 n-chimaerin AA799531 ESTs, Weakly similar to M18.3.p[Caenorhabditis elegans] Common genes downregulated from baseline to 2 hafter acute stress L19699 Rat GTP-binding protein (ral B) AI63933698.51% zinc finger protein 326 Putative Ortholog (mouse) U04998Phosphacan AA892549 ESTs AA893664 TEMO (novel protein; PA200 a putativehuman ortholog) Common genes downregulated from baseline to 24 h afteracute stress M18416 Nerve growth factor-induced (NGFI-A, Egr-1) U75397Krox-24 (NGFI-A, Egr-1) AF023087 Nerve growth factor induced factor A(Egr-1, Krox-24) AB003726 Vesl Genes upregulated in the normal controlsfrom baseline to 30 min after acute stress X56325 2-alpha-1 globin geneAF020618 Progression elevated gene 3 X54686 pJunB AA800680 ESTs, Weaklysimilar to S68418 protein phosphatase 1M chain M110 isoform AA800881ESTs, Weakly similar to SUDY AI178971 Hemoglobin, alpha 1 AA875444 ESTs,similar to dihydropyriminidase-like 2 AA800245 ESTs, Weakly similar toT09013 RING finger protein Fxy - mouse Genes upregulated in the normalcontrols from baseline to 2 h after acute stress AA891127 NucleolinL01624 Serum and glucocorticoid-regulated kinase (sgk) U33500 Retinoldehydrogenase type II AI176456 ESTs, Highly similar toMETALLOTHIONEIN-II C07012 Peptidylpropyl isomerase C-associated proteinAI639415 ESTs, Weakly similar to KERATIN, TYPE I CYTOSKELETAL 21 D16102ATP-stimulated glucocorticoid-receptor translocation promoter U69272Interleukin-15 AA800881 ESTs, Weakly similar to SUDY Genes upregulatedin the normal controls from baseline to 24 h after acute stress AA89316493.82% expressed sequence AA409659 Putative Ortholog (mouse) X94246Pax-8 protein U16253 Corticotropin-releasing factor receptor subtype 2(CRF2R) X14674 Protamine 2 AA800881 ESTs, Weakly similar to SUDYAA799745 CDK5 activator-binding protein C53 U33500 Retinol dehydrogenasetype II AA800738 ESTs, Highly similar to TI60_HUMAN 60 KDA TAT Genesdownregulated in the normal controls from baseline to 30 min after acutestress X67250 R. norvegicus mRNA for n-chimaerin AA799531 ESTs, Weaklysimilar to M18.3.p [Caenorhabditis elegans] AA893596 ESTs, Weaklysimilar to T43458 hypothetical DKFZp434F0621.1 protein Genesdownregulated in the normal controls from baseline to 2 h after acutestress L19699 Rat GTP-binding protein (ral B) AI639336 98.51% zincfinger protein 326 Putative Ortholog (mouse) U04998 Phosphacan AA892549ESTs AA893664 TEMO (novel protein) Genes downregulated in the normalcontrols from baseline to 24 h after acute stress AF036335 NonO/p54nrbhomolog AF063447 Nuclear RNA helicase AF093267 Homer-1b J05592 Proteinphosphatase inhibitor-1 protein M15191 Beta-tachykinin, substance PM32754cd Inhibin alpha-subunit AA818983 Diacylglycerol kinase 90 kDa(Dagk) AA875032 ESTs, hypothetical protein FLJ23306 putative orthologAA892775 Lysozyme (lyz) AA892942 ESTs AI136540 ESTs, Highly similar toTRT3 RAT TROPONIN T, AI145931UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (uae1)AI176710 nuclear receptor subfamily 4, group A, member 3 (Nr4a3) S46131dopamine D1 receptor {promoter} U10071 CART protein X95466 CPG2 protein

TABLE 6 Genes constituting the “abnormal stress response” after exposureto acute stress. Accession Description Genes upregulated in theprenatally stressed from baseline to 30 min after acute stress L26292FSH-regulated protein mRNA M23697 Tissue-type plasminogen activator(t-PA) X06769cds c-fos D28557 RYB-a AF003835 Isopentenyldiphosphate-dimethylallyl diphosphate isomerase X76489cds CD9 Cellsurface glycoprotein AF030091 Activity and neurotransmitter-inducedearly gene 6 (ania-6) AF030088 Activity and neurotransmitter-inducedearly gene 3 (ania-3) M24604 Proliferating cell nuclear antigen(PCNA/cyclin) X58294 Carbonic anhydrase II E03358cds Proteasome(prosome, macropain) subunit, alpha type 2 AA894174 ESTs, Highly similarto ELECTRON TRANSFER FLAVOPROTEIN ALPHA-SUBUNIT PRECURSOR U42627 Dualspecificity phosphatase 6 U38180 Reduced folate carrier membraneglycoprotein U90829 APP-binding protein 1 M62388 Ubiquitin conjugatingenzyme M12112 Angiotensinogen M94918 Beta-globin gene, exons 1-3AF026529 Stathmin-like-protein splice variant RB3 Genes upregulated inthe prenatally stressed from baseline to 2 h after acute stress M93273Somatostatin-receptor type 2 AI176710 Nuclear receptor subfamily 4,group A, member 3 (Nr4a3) X01785 Rat thymocyte mRNA for cell surfaceprotein (MRC OX-2) X76489cds CD9 Cell surface glycoprotein Genesupregulated in the prenatally stressed from baseline to 24 h after acutestress U20796 Nuclear receptor Rev-ErbA-beta/Nr1d2 AF078779 Voltagegated channel like 1 X76489cds CD9 cell surface glycoprotein AA89250086.89% unc-51-like kinase 2 (C. elegans) AA799721 ESTs, highly similarto cysteine and histidine rich 1 M31809 Calcineurin A-beta AA87498297.15% importin beta Putative Ortholog (mouse) AF021350 Natural killercell protein group 2-A (NKG2A) AI175959 Avian sarcoma virus 17 (v-jun)oncogene homolog (Jun) S67769 Solute carrier family 8 (sodium/calciumexchanger), member 1 AB003992 SNAP-25B S81478 PTPase = oxidativestress-inducible protein tyrosine phosphatase U42719 C4 complementprotein Genes downregulated in the prenatally stressed from baseline to30 min after acute stress X16481 parathymosin AF064868 Brain-enrichedguanylate kinase-associated protein 1, Begain C07012 Peptidylpropylisomerase C-associated protein AI639023 89.32% ESTs, Weakly similar toPSF_HUMAN PTB- ASSOCIATEDSPLICING FACTOR [H. sapiens] X58865mRNA PFK-LmRNA for liver phosphofructokinase AA799681 ESTs M88751 Calcium channelbeta subunit-III S70803 Clone p10.15 product [rats, osteosarcomaROS17/2.8, mRNA, 737 nt] AA800942 Complement component 4 (C4a) AI639195ESTs AA893749 EST, Weakly similar to 2206317A protein SS AA893924 ESTs,Highly similar to KLFD_MOUSE Krueppel-like factor 13 U70270UTR#1 Mud-4mRNA Y13714 Osteonectin Genes downregulated in the prenatally stressedfrom baseline to 2 h after acute stress L08228exon#22N-methyl-D-aspartate receptor (NMDAR1) gene C07012 Peptidylprolylisomerase C-associated protein X58865mRNA PFK-L mRNA for liverphosphofructokinase X17012mRNA IGFII gene for insulin-like growth factorII AA893924 ESTs, Highly similar to KLFD_MOUSE Krueppel-like factor 13M62752 Statin-like protein AA892053 ESTs, Highly similar to T42204chromatin structural protein homolog Supt5hp —mouse [M. musculus]AF093536 Beta defensin-1 (BD-1) mRNA U70270UTR#1 Mud-4 AF065433 Rattusnorvegicus Bcl-2 related ovarian death gene product Bim/BOD AI172476TGFB inducible early growth response (Tieg) AF064868 Brain-enrichedguanylate kinase-associated protein 1 (BEGAIN) X60469mRNA FE65 adaptorprotein AI639023 89.32% ESTs, Weakly similar to PSF_HUMAN PTB-ASSOCIATEDSPLICING FACTOR L40364 MHC class I RT1.O type - 149 processed pseudogeneD12927 Transcription elongation factor S-II AI014135 Beta-carotene 15,15′-dioxygenase (Bcdo) Genes downregulated in the prenatally stressedfrom baseline to 24 h after acute stress AF065433 Bcl-2 related ovariandeath gene product Bim/BOD AF093536 Beta defensin-1 (BD-1) D86711 Zincfinger, DHHC domain (putative ortholog) M93257Cathechol-O-methyltransferase AA800602 Chemokine (C—X3—C motif) ligand1, neurotactin AA874843 CD36 antigen (collagen type I receptor,thrombospondin receptor)-like 1 U49099 cis-Golgi p28 (p28) U65656 Matrixmetalloproteinase 2 X16481 Parathymosin

TABLE 7 Genes significantly differentially expressed at individualtimepoints following acute stress in the prenatally non-stressed (NS)_and prenatally stressed PNS frontal pole (“abnormal stress response”).Accession Description Genes upregulated in the PNS group at baselineU66707 Densin-180 AA900582 Alpha-2-macroglobulin AI012942 cell divisioncycle 25B (cdc25b) AF064868 Begain AI044716 Neuronal pentraxin precursorAF068136 Rattus norvegicus G alpha interacting protein (GAIP) X58865PFK-L mRNA for liver phosphofructokinase AI179150 Cytochrome b oxidaseAA800881 cDNA K02248 Somatostatin-14 gene AF001423 N-methyl-D-aspartatereceptor NMDAR2A subunit Genes downregulated in the PNS group atbaseline X01785 Rat thymocyte mRNA for cell surface protein (MRC OX-2)AA892376 93.22% protein associated with PRK1 Putative Ortholog (highlyconserved) AA866459 ESTs, Highly similar to hypothetical protein MGC4175[Homo sapiens] AA892483 86.36% KIAA0877 protein Putative OrthologAA891969 89.35% nuclear DNA-binding protein Putative Ortholog AJ001641Brain-1 (Brn-1) AA799636 91.33% ESTs, Highly similar to T00362hypothetical protein KIAA0675 AA945054 cytochrome b5 (Cyb5) AI008131S-Adenosylmethionine decarboxylase 1A AA893670 ESTs AI227715Retinoblastoma-related gene (Rb2) AA799576 ESTs, Highly similar toT46259 hypothetical protein DKFZp761E0323.1 AA799791 ESTs, Weaklysimilar to T34021 protein kinase SK2 - rat AA874982 97.15% importin betaPutative Ortholog X76489cds RNCD9 CD9 mRNA for cell surface glycoproteinY09000 Dendrin AA859663 ESTs U77931 Ribin AI145367 Adenylylcyclase-associated protein 2 (Cap2) D10666 Neural visinin-like protein(NVP) D38560 CyclinG-associated kinase (Gak) AB003992 SNAP-25B AA85952097.45% Homo sapiens cDNA FLJ31057 fis, clone HSYRA2000787 AB003991SNAP-25A AA894264 ESTs Genes upregulated in the PNS group 30 min afteracute stress AF068136 G alpha interacting protein (GAIP) AI639157 cDNAclone rx00682 3 AA891901 Polypyrimidine tract binding protein M30691Ly6-C antigen AI234950 Acid phosphatase 2, lysozymal U66707 Densin-180Genes upregulated in the PNS group 2 h after acute stress M30691 Ly6-Cantigen X57514 GABA(A) receptor gamma-1 subunit S79523 Selectin,lymphocyte membrane protein A.11 (Sell) AI639165 cDNA clone rx01762 3D00913 Intercellular adhesion molecule-1 AJ224680 Cyclicnucleotide-gated channel beta subunit 1 (Cngp1) AA893711 ESTs, Weaklysimilar to DnaJ (Hsp40) homolog, subfamily B, member 9; Microvascularendothelial differentiation gene 1 AI639422 ESTs, Moderately similar toCAQC_RAT CALSEQUESTRIN D14015 Cyclin E L31840 Nuclear pore complexprotein NUP107 AF087674 Insulin receptor substrate 2 (IRS-2) X05472 2.4kb repeat DNA right terminal region Genes downregulated in the PNS group2 h after acute stress M62752 Statin-related protein (s1) gene X07729Neuron-specific enolase, exons 8-12 AA955388 ATPase isoform 2, Na+K+transporting, beta polypeptide 2 AA799479 ESTs, Moderately similar toNUIM_HUMAN NADH-ubiquinone oxidoreductase 23 kDa subunit, mitochondrialprecursor (Complex I- 23 KD) (CI-23 KD) (TYKY subunit) H32977 EST108553Rattus norvegicus cDNA X54531 Dynamin-1 AA799791 ESTs, Weakly similar toT34021 protein kinase SK2 M64986 Amphoterin Genes upregulated in the PNSgroup 24 h after acute stress AI145494 Synapsin-2 U66707 Densin-180U77583 Casein kinase I alpha L (CKIaL) K02248 Somatostatin-14 geneAA892797 ESTs, Highly similar to A33792 phosphoglycerate kinase (EC2.7.2.3) - rat U70268 mud-7 M88751 Calcium channel beta subunit-IIIGenes downregulated in the PNS group 24 h after acute stress AA893164ESTs AI227715 Retinoblastoma-related gene Rb2 M35300 Pancreaticsecretory trypsin inhibitor-like protein (PSTI) AB016160 GABAB receptor1c, complete cds AI101103 Vamp2 U77931 Ribin AI145367 Adenylylcyclase-associated protein 2 Cap2 Y09000 Dendrin

1-26. (canceled)
 27. A method for screening a subject for schizophrenia,bipolar disorder and/or ADHD or a subject at risk of developingschizophrenia, bipolar disorder and/or ADHD, comprising either: A)detecting a level of expression of at least one gene, selected from thegroup consisting of (Table 1), in a sample of frontal pole tissueobtained from the subject to provide a first value, with the provisothat if expression of only one gene is detected that the gene is not anyof the genes selected from the group consisting of (Table 3); andcomparing the first value with a level of expression of the at least onegene in said detecting step in a sample of frontal pole tissue obtainedfrom a disease-free subject, wherein a greater expression level in thesubject sample compared to the sample from the disease-free subject ofat least one gene with accession number X57514, U66707, AI639165,AF064868, or AI145494 or wherein a lower expression level in the subjectsample compared to the sample from the disease-free subject of at leastone gene with accession number U20643, H31232, S61973, AB016160, X85184,J04063, S81353, M33025, AA858621, M74494, AA866358, M64986, AI227715,U77931, AA89392, AA859633, AA891969, AI145367, AA893711, Y09000,AA891940, D26564, AI101103, or AF014009 is indicative of the subjecthaving schizophrenia, bipolar disorder and/or ADHD or at risk ofdeveloping schizophrenia, bipolar disorder and/or ADHD; or B) detectinga level of expression of at least one gene, selected from the groupconsisting of (Table 2), in a sample of hypothalamus tissue obtainedfrom the subject to provide a first value, with the proviso that ifexpression of only one gene is detected that the gene is not any of thegenes selected from the group consisting of (Table 3); and comparing thefirst value with a level of expression of the at least one gene in saiddetecting step in a sample of hypothalamus tissue obtained from adisease-free subject, wherein a greater expression level in the subjectsample compared to the sample from the disease-free subject of at leastone gene with accession number AA946532, AB016160, U57500, L17127,M28648, L24776, AB020504, U77931, U75927, AF096269, AA892801, U49099,AA859597, AA875427, AF028784, L22760, AI014135, AI104513, AA894148,AI073204 or D38468, wherein a lower expression level in the subjectsample compared to the sample from the disease-free subject of at leastone gene with accession number M13100, M59980, M18331, M36418, X57764,U62897, S71570, M16112, AI008131, X12744, AI230260, U48245, U66707,H31692, D89863, X06564, M91234, U31554, AA875659, AA799421, AI014091,AB012234, U09793, AA800513, AI639381, AA799515, X83546, AI013194,AB008538, X52817, AA893065, H31588, AA859832, AI008074, AA851749,AA894321, AI227608, E13644, AA925762, M72422, M17526, U45479, U86635,AA894089, or AA891069 is indicative of the subject having schizophrenia,bipolar disorder and/or ADHD or at risk of developing schizophrenia,bipolar disorder and/or ADHD; or C) detecting a level of expression ofat least one gene, selected from the group consisting of (Table 7), in asample of frontal pole tissue obtained from the subject to provide afirst value, with the proviso that if expression of only one gene isdetected that the gene is not any of the genes selected from the groupconsisting of (Table 3); and comparing the first value with a level ofexpression of the at least one gene in said detecting step in a sampleof frontal pole tissue obtained from a disease-free subject, wherein agreater expression level in the subject sample compared to the samplefrom the disease-free subject of at least one gene with accession numberU66707, AA900582, AI012942, AF064868, AI044716, AF068136, X58865,AI179150, AA800881, K02248, AF001423, AI639157, AA891901, M30691,AI234950, X57514, S79523, AI639165, D00913, AJ224680, AA893711,AI639422, D14015, L31840, AF087674, X05472, AI145494, U77583, AA892797,U70268 or M88751 or wherein a lower expression level in the subjectsample compared to the sample from the disease-free subject of at leastone gene with accession number AA892376, AA866459, AA892483, AA891969,AJ001 641, AA799636, AA945054, AI008131, AA893670, AI227715, AA799576,AA799791, AA874982, X76489cds, Y09000, AA859663, U77931, AI145367,D10666, D38560, AB003992, AA859520, AB003991, AA894264, AA955388,X07729, M62752, AA799479, H32977, X54531, M64986, AA893164, M35300,AB016160 or AI101103 is indicative of the subject having schizophrenia,bipolar disorder and/or ADHD or at risk of developing schizophrenia,bipolar disorder and/or ADHD.
 28. The method of claim 27, wherein thelevel of expression of at least two genes, selected from the groupconsisting of (Tables 1, 2 and 7), are detected.
 29. The method of claim27 (A) or (B), wherein the at least one gene is selected from the groupconsisting of genes with accession numbers: U20643, H31232, S61973,AB016160, X85184, J04063, S81353, M33025, X57514, AA858621, M74494,M13100, M59980, AA946532, M18331, M36418, X57764, U62897, U57500,L17127, S71570, M16112, AI008131, M28648, X12744, AI230260, U86635, andD38468.
 30. The method of claim 27, wherein the level of expression ofthe gene is determined by detecting the level of expression of a mRNAcorresponding to the gene.
 31. The method of claim 30, wherein the levelof expression of mRNA is detected by techniques selected from the groupconsisting of Microarray analysis, Northern blot analysis, reversetranscription PCR, and real time quantitative PCR.
 32. The method ofclaim 27, wherein the level of expression of the gene is determined bydetecting the level of expression of a protein encoded by the gene. 33.A method for monitoring the progression of schizophrenia, bipolardisorder, and/or ADHD in a subject having, or at risk of having,schizophrenia, bipolar disorder, and/or ADHD, comprising either: A)measuring a level of expression of at least one gene, selected from thegroup consisting of (Table 1), over time in the frontal pole tissuesample obtained from the subject with the proviso that if expression ofonly one gene is detected that the gene is not any of the genes selectedfrom the group consisting of (Table 3), wherein an increase in the levelof expression of the at least one gene with accession number: X57514,U66707, AI639165, AF064868, or AI145494 over time is indicative of theprogression of schizophrenia, bipolar disorder, and/or ADHD in thesubject, or wherein a decrease in the level of expression of the atleast one gene with accession number U20643, H31232, S61973, AB016160,X85184, J04063, S81353, M33025, AA858621, M74494, AA866358, M64986,AI227715, U77931, AA89392, AA859633, AA891969, AI145367, AA893711,Y09000, AA891940, D26564, AI01103, or AF014009 over time is indicativeof the progression of schizophrenia, bipolar disorder, and/or ADHD inthe subject; or B) measuring a level of expression of at least one gene,selected from the group consisting of (Table 2), over time in thehypothalamus tissue sample obtained from the subject with the provisothat if expression of only one gene is detected that the gene is not anyof the genes selected from the group consisting of (Table 3), wherein anincrease in the level of expression of the at least one gene withaccession number AA946532, AB016160, U57500, L17127, M28648, L24776,AB020504, U77931, U75927, AF096269, AA892801, U49099, AA859597, M875427,AF028784, L22760, AI014135, AI104513, M894148, AI073204 or D38468 overtime is indicative of the progression of schizophrenia, bipolardisorder, and/or ADHD in the subject, or wherein a decrease in the levelof expression of the at least one gene with accession number M13100,M59980, M18331, M36418, X57764, U62897, S71570, M16112, AI008131,X12744, AI230260, U48245, U66707, H31692, D89863, X06564, M91234,U31554, M875659, AA799421, AI014091, AB012234, U09793, M800513,AI639381, AA799515, X83546, AI013194, AB008538, X52817, AA893065,H31588, AA859832, AI008074, AA851749, AA894321, AI227608, E13644,AA925762, M72422, M17526, U45479, U86635, AA894089, or AA891069 overtime is indicative of the progression of schizophrenia, bipolardisorder, and/or ADHD in the subject.
 34. The method of claim 33,wherein the level of expression of at least two genes, selected from thegroup consisting of (Table 1 and 2), are measured.
 35. The method ofclaim 33, wherein the at least one gene is selected from the groupconsisting of the genes with accession number U20643, H31232, S61973,AB016160, X85184, J04063, S81353, M33825, X57514, AA858621, M74494,M13100, M59980, AA946532, M18331, M36418, X57764, U62897, U57500,L17127, S71570, M16112, AI008131, M28648, X12744, AI230260, U86635, andD38468.
 36. A method for identifying agents for use in the treatment ofschizophrenia, bipolar disorder, and for ADHD, comprising either: A. a)contacting a sample of cells expressing at least one gene selected fromthe group consisting of (Tables 1 and 2) with a candidate agent; b)detecting a level of expression of at least one gene in said cells, withthe proviso that if expression of only one gene is detected that thegene is not any of the genes selected from the group consisting of(Table 3); and c) comparing the level of expression of the at least onegene in the sample in the presence of the candidate agent with a levelof expression of the at least one gene in cells that are not contactedwith the candidate agent, wherein for genes with accession numberU20643, H31232, S61973, X85184, J04063, S81353, M33025, AA858621,M74494, AA866358, M64986, AI227715, AA89392, AA859633, AA891969,AI145367, AA893711, Y09000, AA891940, D26564, AI101103, AF014009,M13100, M59980, M18331, M36418, X57764, U62897, S71570, M16112,AI008131, X12744, AI230260, U48245, H31692, D89863, X06564, M91234,U31554, AA875659, AA799421, AI014091, AB012234, U09793, AA800513,AI639381, AA799515, X83546, AI013194, AB008538, X52817, AA893065,H31588, AA859832, AI008074, AA851749, AA894321, AI227608, E13644,AA925762, M72422, M17526, U45479, U86635, AA894089, or AA891069 anincreased level of expression of the at least one gene in the sample inthe presence of the candidate agent relative to the level of expressionof the at least one gene in the sample in the absence of the candidateagent, or wherein for genes with accession number X57514, AI639165,AF064868, AI145494, AA946532, U57500, L17127, M28648, L24776, AB020504,U75927, AF096269, AA892801, U49099, AA859597, AA875427, AF028784,L22760, AI014135, AI104513, AA894148, AI073204 or D38468 a decreasedlevel of expression of the at least one gene in the sample in thepresence of the candidate agent relative to the level of expression ofthe at least one gene in the sample in the absence of the candidateagent and for genes with accession numbers AB016160 and U77931 anincreased level of expression with respect to the frontal pole, and adecreased expression with respect to the hypothalamus and for a genewith accession number U66707 an increased expression with respect tohypothalamus and an decreased expression with respect to the frontalpole is indicative of an agent useful in the treatment of schizophrenia,bipolar disorder, and/or ADHD; or B. a) contacting a sample of cellsexpressing at least one gene, selected from the group consisting of(Table 7) with a candidate agent; b) detecting level of expression of atleast one gene in said cells with the proviso that if expression of onlyone gene is detected that the gene is not any of the genes selected fromthe group consisting of (Table 3); and c) comparing the level ofexpression of the at least one gene in the sample in the presence of thecandidate agent with a level of expression of the at least one gene incells that are not contacted with the candidate agent, wherein for geneswith accession number: AA892376, AA866459, AA892483, AA891969, AJ001641,AA799636, AA945054, AI008131, AA893670, AI227715, AA799576, AA799791,AA874982, X76489cds, Y09000, AA859663, U77931, AI145367, D10666, D38560,AB003992, AA859520, AB003991, AA894264, AA955388, X07729, M62752,AA799479, H32977, X54531, M64986, AA893164, M35300, AB016160 or AI101103an increased level of expression of the at least one gene in the samplein the presence of the candidate agent relative to the level ofexpression of the at least one gene in the sample in the absence of thecandidate agent: or wherein for genes with accession number: U66707,AA900582, AI012942, AF064868, AI044716, AF068136, X58865, AI79150,AA800881, K02248, AF001423, AI639157, AA891901, M30691, AI234950,X57514, S79523, AI639165, D00913, AJ224680, AA893711, AI639422, D14015,L31840, AF087674, X05472, AI145494, U77583, AA892797, U70268 or M88751 adecreased level of expression of the at least one gene in the sample inthe presence of the candidate agent relative to the level of expressionof the at least one gene in the sample in the absence of the candidateagent; is indicative of an agent useful in the treatment ofschizophrenia, bipolar disorder, and/or ADHD.
 37. The method of claim36, wherein the level of expression of at least two genes in the sampleis detected the detecting step (b).
 38. The method of claim 36(A),wherein the at least one gene is selected from the group consisting ofgenes with accession number U20643, H31232, S61973, AB016160, X85184,J04063, S81353, M33025, X57514, AA858621, M74494, M13100, M59980,AA946532, M18331, M36418, X57764, U62897, U57500, L17127, S71570,M16112, AI008131, M28648, X12744, AI230260, U86635, and D38468.
 39. Amethod of treating and preventing schizophrenia, bipolar disorder,and/or ADHD in patient in need thereof, comprising either: A)administering to said patient an effective amount of an agent that caninduce a decrease in the expression of at least one gene with accessionnumber U66707, AA900582, AI012942, AF064868, AI044716, AF068136, X58865,AI179150, AA800881, K02248, AF001423, AI639157, AA891901, M30691,AI234950, X57514, S79523, AI639165, D00913, AJ224680, AA893711,AI639422, D14015, L31840, AF087674, X05472, AI145494, U77583, AA892797,U70268 and/or M88751; and/or induce an increase of at least one genewith accession number AA892376, AA866459, AA892483, AA891969, AJ001641,AA799636, AA945054, AI008131, AA893670, AI227715, AA799576, AA799791,AA874982, X76489cds, Y09000, AA859663, U77931, AI145367, D10666, D38560,AB003992, AA859520, AB003991, AA894264, AA955388, X07729, M62752,AA799479, H32977, X54531, M64986, AA893164, M35300, AB016160 and/or forAI101103, with the proviso that if expression of only one gene isaltered that the gene is not any selected from the group consisting of(Table 3); or B) administering to said patient an effective amount of anagent that can induce a decrease in the expression of at least one genewith accession number X57514, AI639165, AF064868, AI145494, AA946532,U57500, L17127, L24776, A8020504, U75927, AF096269, AA892801, U49099,AA859597, AA875427, AF028784, L22760, AI014135, AI104513, AA894148,AI073204 and/or D38468; and/or induce an increase of at least one genewith accession number U20643, H31232, S61973, X85184, J04063, S81353,M33025, AA858621, M74494, AA866358, M64986, AI227715, AA89392, AA859633,AA891969, AI145367, AA893711, Y09000, AA891940, D26564, AI101103,M28648, AF014009, M13100, M59980, M18331, M36418, X57764, U62897,S71570, M16112, AI008131, X12744, AI230260, U48245, H31692, D89863,X06564, M91234, U31554, AA875659, AA799421, AI014091, AB012234, U09793,AA800513, AI639381, AA799515, X83546, AI03194, AB008538, X52817,AA893065, H31588, AA859832, AI008074, AA851749, AA894321, AI227608,E13644, AA925762, M72422, M17526, U45479, U86635, AA894089, and/orAA891069; and/or for genes AB016160 and/or U7793-lan increase withrespect to frontal pole expression and a decrease to the hypothalamusexpression; and/or for the gene U66707 a decrease with respect to thefrontal pole and an increase with respect to the hypothalamus, with theproviso that if expression of only one gene is altered that the gene isnot any gene of selected from the group consisting of (Table 3).
 40. Amethod for monitoring the efficacy of a treatment of a subject havingschizophrenia, bipolar disorder, and/or ADHD or at risk of developingschizophrenia, bipolar disorder, and/or ADHD with an agent, comprisingeither: A. a) obtaining a pre-administration sample in the frontal lobefrom the subject prior to administration of the agent; b) detecting alevel of expression of at least one gene, selected from the groupconsisting of (Tables 1 and 7), in the pre-administration sample, withthe proviso that if expression of only one gene is detected that thegene is not any gene selected from the group consisting of (Table 3); c)obtaining one or more post-administration samples from the subject; d)detecting a level of expression of the at least one gene in thepost-administration sample or samples; e) comparing the level ofexpression of the at least one gene in the pre-administration samplewith the level of expression of the at least one gene in thepost-administration sample; and f) adjusting the administration of theagent accordingly; or B. a) obtaining a pre-administration sample in thehypothalamus from the subject prior to administration of the agent; b)detecting a level of expression of at least one gene, selected from thegroup consisting of (Table 2), in the pre-administration sample, withthe proviso that if expression of only one gene is detected that thegene is not any gene selected from the group consisting of (Table 3); c)obtaining one or more post-administration samples from the subject; d)detecting a level of expression of the at least one gene in thepost-administration sample or samples; e) comparing the level ofexpression of the at least one gene in the pre-administration samplewith the level of expression of the at least one gene in thepost-administration sample; and f) adjusting the administration of theagent accordingly.
 41. The method of claim 40, wherein the level ofexpression of at least two genes, selected from the group consisting of(Tables 1, 2 and 7), is detected in step (b).
 42. The method of claim40, wherein the level of expression of mRNA is detected by techniquesselected from the group consisting of Microarray analysis, Northern blotanalysis, reverse transcription PCR and real time quantitative PCR. 43.A transgenic mouse whose genome comprises a disruption of any of theendogenous genes selected from the group consisting of (Table 1, 2 and7), wherein said disruption comprises the insertion of a transgene, andwherein said disruption results in said transgenic mouse not exhibitingnormal expression of any of said endogenous genes with the proviso thatif only one gene is disrupted it is not one of the genes of selectedfrom the group consisting of (Table 3).
 44. A method for producing arodent having schizophrenia, bipolar disorder, and/or ADHD or at risk ofdeveloping schizophrenia, bipolar disorder, and/or ADHD, comprisingsubjecting the rodent to variable repeated stress during the last weekof the gestation, starting on embryonic day 14 and continuing until thenatural delivery of a pup at about embryonic day 22.